RU2789884C2 - Antibodies to myostatin, polypeptides containing fc-region variants, and their application methods - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology, in particular to an antibody, which specifically binds to latent myostatin, as well as to its production method. Nucleic acid encoding the above-mentioned antibody, as well as a cell containing it are also disclosed.

EFFECT: invention is effective for specific binding to myostatin.

7 cl, 31 dwg, 30 tbl, 34 ex

Description

The field of technology to which the invention belongs

The present invention relates to antibodies to myostatin and method of their use. The present invention also relates to polypeptides containing variants of the Fc regions, and methods of their use.

Prerequisites for the creation of the invention

Myostatin, also referred to as differentiation factor-8 (GDF8), is a secreted protein and is a member of the transforming growth factor beta (TGF-beta) protein superfamily. Members of this superfamily have growth regulating and morphogenetic properties (see, for example, NPL 1, NPL 2 and PTL 1). Myostatin is expressed primarily in developing adult skeletal and musculature and functions as a negative regulator of muscle growth. Systemic overexpression of myostatin in adult mice leads to muscle wasting (see, for example, NPL 3), while in the opposite case, mice with "turned off" myostatin are characterized by hypertrophy and hyperplasia of skeletal muscles, leading to a 2-3-fold increase in muscle mass compared to their wild-type littermates (see, for example, NPL 4).

Like other members of the TGF-beta family, myostatin is synthesized as a large precursor protein that contains an N-terminal propeptide domain and a C-terminal domain, which is an active molecule (see, for example, NPL 5; PTL 2). The two myostatin precursor molecules are covalently linked via a single disulfide bond present in the C-terminal domain of the growth factor. Active mature myostatin (a disulfide-linked homodimer consisting of the C-terminal growth factor domain) is released from the myostatin precursor through several steps of proteolytic processing. At the first stage of the myostatin activation pathway, the peptide bond between the N-terminal propeptide domain and the C-terminal domain of the growth factor is cleaved; Arg266-Asp267 using a furin-type proprotein convertase in both strands of the homodimeric precursor. However, the resulting three peptides (two propeptides and one mature myostatin (i.e., a disulfide-linked homodimer consisting of growth factor domains)) remain associated, forming a non-covalent inactive complex, which is referred to as "latent myostatin". Mature myostatin can then be released from latent myostatin via propeptide cleavage. Representatives of the family of metalloproteinases, which are bone morphogenetic protein (protein involved in osteogenesis) 1 (BMP1), cleave a single peptide bond in the propeptide, Arg98-Asp99, which is accompanied by the release of mature active myostatin in the form of a homodimer (see, for example, NPL 6) . In addition, latent myostatin can also be activated in vitro by dissociating the complex with either acid treatment or heat treatment (see, for example, NPL 7).

Myostatin exerts its actions through a family of transmembrane heterotetrameric serine/threonine kinase receptors, the activation of which increases receptor transphosphorylation, leading to stimulation of serine/threonine kinase activity. It has been established that the myostatin pathway includes an active myostatin dimer that binds to the activin-type HB receptor (ActRIIB) with high affinity, which further leads to the emergence and activation of transphosphorylation of the low-affinity receptor, activin-like kinase 4 (ALK4) or activin-like kinase 5 (ALK5). It was also found that the Smad 2 and Smad 3 proteins are then activated and form a complex with Smad 4, which then translocates into the nucleus to activate the transcription of the target gene. It has been demonstrated that ActRIIB has the ability to mediate the effects of myostatin in vivo, since the expression of the dominant-negative form of ActRIIB in mice mimics myostatin gene shutdown (see, for example, NPL 8).

A variety of disorders or conditions are associated with muscle wasting (i.e., loss or functional impairment of muscle tissue), such as muscular dystrophy (MD; including Duchenne muscular dystrophy), amyotrophic lateral sclerosis (ALS), muscle atrophy, organ atrophy, infirmity, congestive obstructive pulmonary disease (COPD), sarcopenia and cachexia resulting from cancer or other disorders, as well as kidney disease, heart failure, or liver disease and disease. Patients may benefit from increased muscle mass and/or muscle strength; however, only limited treatments for these disorders are currently available. Thus, because of its role as a negative regulator of skeletal muscle growth, myostatin becomes an important target for therapeutic or prophylactic intervention in said disorders or conditions, or for monitoring the development of said disorders or conditions. In particular, agents that inhibit the activity of myostatin may be valuable therapeutic agents.

Inhibition of myostatin expression results in both muscle hypertrophy and hyperplasia (NPL 4). Myostatin is a negative regulator of muscle regeneration after injury, and the absence of myostatin in myostatin-null mice results in accelerated muscle regeneration (see, for example, NPL 9). Anti-myostatin antibodies (GDF8), which are described, for example, in PTL 3, PTL 4, PTL 5, PTL 6 and PTL 7, and PTL 8, PTL 9 and PTL 10, have been found to bind to myostatin and inhibit myostatin activity in vitro. and in vivo, including myostatin activity associated with negative regulation of skeletal muscle mass. Myostatin-neutralizing antibodies increase body weight, skeletal muscle mass and muscle size, as well as skeletal muscle strength in wild-type mice (see, for example, NPL 10) and in mdx mice (muscular dystrophy model) (see, for example, NPL 11 ; NPL 12). However, the antibodies known in the art are all specific for mature myostatin but not for latent myostatin, and the described strategies for inhibiting myostatin activity have used antibodies that can bind to and neutralize mature myostatin.

Antibodies are attracting attention as pharmaceuticals due to their high blood stability and few side effects (see, for example, NPL 13 and NPL 14). Nearly all current commercially available therapeutic antibodies are of the human IgG1 subclass. One of the known functions of IgG-class antibodies is antibody-mediated cell-dependent cytotoxicity (hereinafter referred to as ADCC activity in the context of the present description) (see, for example, NPL 15). For an antibody to exhibit ADCC activity, the Fc region of an antibody must bind to the Fc-gamma receptor (hereinafter referred to as Fc-gamma R), which is an antibody-binding receptor present on the surface of effector cells such as killer cells, natural killer cells and activated macrophages.

For humans, the isoforms Fc-gamma RIa (CD64A), Fc-gamma RIIa (CD32A), Fc-gamma RIIb (CD32B), Fc-gamma RIIIa (CD 16A) and Fc-gamma RIIIb (CD16B) are described as proteins of the Fc- gamma R, in addition, the corresponding allotypes are also described (see, for example, NPL 16). Fc-gamma RIa, Fc-gamma RIIa and Fc-gamma RIIIa are called activating Fc-gamma R because they have immunologically active functions, and Fc-gamma RIIb is called inhibitory Fc-gamma R because it has immunosuppressive functions (see, e.g., , NPL 17).

Several amino acid residues in the antibody hinge region and the CH2 domain and a sugar chain attached to Asn at position 297 (EU numbering) that is linked to the CH2 domain (see ., for example, NPL 18, NPL 19 and NPL 20). To date, various variants have been obtained that have the ability to bind to Fc-gamma R, primarily antibodies with mutations introduced at these sites; and Fc region variants having higher binding activities for the activating Fc-gamma R (see, for example, PTL 11, PTL 12, PTL 13 and PTL 14).

When the activating Fc-gamma R cross-links with the immune complex, it phosphorylates immunoreceptor tyrosine-based activating motifs (ITAM) within the intracellular domain or common gamma chain of the FcR (interaction partner), activates the SYK signal transducer, and triggers an inflammatory immune response. by initiating activation of the signal cascade (see, for example, NPL 21).

Fc-gamma R11b is the only Fc-gamma R that is expressed on B cells (see, for example, NPL 22). It has been established that the interaction of the Fc region of the antibody with the Fc-gamma of RIIb suppresses the primary immune response of B cells (see, for example, NPL 23). In addition, it has been described that when Fc-gamma RIIb on B cells and the B cell receptor (BCR) are cross-linked via an immune complex in the blood, B cell activation and antibody production by B cells is suppressed (see e.g. , NPL 24). This immunosuppressive signal transduction mediated by BCR and Fc-gamma RIIb requires an immunoreceptor tyrosine-based inhibitory motif (ITIM) within the intracellular domain of Fc-gamma RIIb (see, for example, NPL 25 and NPL 26). When ITIM is phosphorylated after signal transduction, SH2-containing inositol polyphosphate 5-phosphatase (SHIP) is recruited, transduction of the other Fc-gamma R signaling cascade is inhibited, and the inflammatory immune response is suppressed (see, for example, NPL 27). In addition, only Fc-gamma RIIb aggregation is known to result in transient suppression of calcium influx due to BCR cross-linking and B-cell proliferation in a BCR-independent manner without inducing apoptosis of IgM-producing B cells (see, e.g., NPL 28 ).

Fc-gamma RIIb is also expressed on dendritic cells, macrophages, activated neutrophils, mast cells, and basophils. Fc-gamma RIIb inhibits activating Fc-gamma R functions such as phagocytosis and release of pro-inflammatory cytokines in these cells and suppresses inflammatory immune responses (see, for example, NPL17).

To date, to study the immunosuppressive functions of the Fc-gamma RIIb carried out studies using mice with "turned off" Fc-gamma RIIb. With their help, it was found that in mice with "off" Fc-gamma RIIb, the humoral immune response is not regulated appropriately (see, for example, NPL 29), sensitivity to collagen-induced arthritis (CIA) increases (see, for example, NPL 30 ), lupus-like symptoms and Goodpasture-like symptoms are present (see, for example, NPL 31).

In addition, human autoimmune diseases are known to be associated with Fc-gamma RIIb dysregulation. For example, a relationship has been described between genetic polymorphisms in the transmembrane region and promoter region of Fc-gamma RIIb and the incidence of systemic lupus erythematosus (SLE) (see, for example, NPL 32, NPL 33, NPL 34, NPL 35 and NPL 36) and reduced expression Fc-gamma RIIb on the surface of B cells in SLE patients (see, for example, NPL 37 and NPL 38).

Based on mouse model data and the clinical results themselves, Fc-gamma RIIb is believed to play a role in the control of autoimmune diseases and inflammatory diseases through recruitment primarily of B cells, and is a promising target molecule for the control of autoimmune diseases. and inflammatory diseases.

It is known that IgG1, which are mainly used as commercially available therapeutic antibodies, not only bind to Fc-gamma RIIb, but also have strong binding to activating Fc-gamma R (see, for example, NPL 39). This makes it possible to develop therapeutic antibodies with greater immunosuppressive properties than those of IgG1 by using an Fc region with increased ability to bind to Fc-gamma RIIb or with improved binding selectivity to Fc-gamma RIIb compared to binding to activating Fc- gamma R. For example, it has been suggested that the use of an antibody having a variable region that binds to BCR and an Fc with increased ability to bind to Fc-gamma RIIb can inhibit B cell activation (see e.g. NPL 40) . Cross-linking of Fc-gamma RIIb on B cells and IgE bound to the B cell receptor has been described to suppress B cell differentiation into plasma cells, resulting in suppression of IgE production; and in mice transplanted with human PBMCs, human IgG and IgM concentrations are maintained while human IgE concentrations are reduced (see, for example, NPL 41). It has been described that when RIIB Fc-gamma and CD79b, which is a constitutive molecule of the B-cell receptor complex, are cross-linked with an antibody, in addition to affecting IgE, B-cell proliferation is suppressed in vitro and arthritis symptoms are alleviated in a collagen-induced arthritis model ( see, for example, NPL 42).

In addition to affecting B cells, cross-linking of Fc-epsilon RI and Fc-gamma RIIb on mast cells using molecules in which an IgG Fc region with increased ability to bind to Fc-gamma RIIb is fused to an IgE Fc region , which binds to the IgE receptor Fc-epsilon RI, causes phosphorylation of Fc-gamma RIIb, thereby suppressing Fc-epsilon RI-dependent calcium influx. This suggests that inhibition of degranulation via Fc-gamma RIIb stimulation can be achieved by increasing Fc-gamma RIIb binding (see, eg, NPL 43).

Thus, an antibody that has an Fc with improved ability to bind to Fc-gamma RIIb is expected to be promising as a therapeutic agent in inflammatory diseases such as autoimmune disease.

In addition, the activation of macrophages and dendritic cells by Toll-like receptor 4 upon stimulation with LPS has been reported to be suppressed in the presence of an antibody-antigen immune complex, and it is suggested that this also represents immune complex actions via Fc-gamma RIIb ( see, for example, NPL 44 and NPL 45). Thus, it is expected that the use of antibodies with increased ability to bind to Fc-gamma RIIb may enhance TLR-mediated inhibitory effects on signal activation; therefore, it is expected that these antibodies may be promising as therapeutic agents in inflammatory diseases such as autoimmune diseases.

In addition, it is expected that mutants with increased ability to bind to Fc-gamma RIIb may be promising as therapeutic agents in cancer, as well as in inflammatory diseases such as autoimmune diseases. To date, it has been established that the Fc-gamma of RIIb plays an important role in the agonistic activity of agonistic antibodies to the TNF receptor superfamily. In particular, it can be assumed that interaction with Fc-gamma RIIb is required for the manifestation of the agonistic activity of antibodies to CD40, DR4, DR5, CD30 and CD137, which are members of the TNF receptor family (see, for example, NPL 46, NPL 47, NPL 48, NPL 49, NPL 50, NPL 51 and NPL 52). NPL 46 demonstrates that the use of antibodies with increased ability to bind to Fc-gamma RIIb increases the antitumor activity of anti-CD40 antibodies. Thus, it can be assumed that antibodies with increased ability to bind to Fc-gamma RIIb may influence the increase in agonist activity of agonist antibodies, including antibodies to the TNF receptor superfamily.

In addition, cell proliferation has been found to be suppressed by using an antibody that recognizes Kit, a type of receptor tyrosine kinase (RTK), to cross-link Fc-gamma RIIb and Kit on Kit-expressing cells. Similar actions are described even in cases in which said Kit was constitutively activated and had mutations that cause tumorigenesis (see, for example, NPL 53). Thus, it can be assumed that the use of antibodies with increased ability to bind to RIIb Fc-gamma may increase the inhibitory effects on cells that express RTK having constitutively activating mutations.

Antibodies having Fc with increased Fc-gamma RIIb-binding activity have been described (see, for example, NPL 40). This publication describes the enhancement of Fc-gamma RIIb binding activity by introducing changes such as S267E/L328F, G236D/S267E and S239D/S267E to the Fc region of an antibody. Among them, the antibody with the introduced S267E/L328F mutation most strongly bound to Fc-gamma RIIb, and it retained the same level of binding to Fc-gamma RIa and Fc-gamma RIIa type H, in which the residue at position 131 Fc-gamma RIIa is a His that is found in naturally occurring IgG1. However, another publication describes that this change increases the binding of Fc-gamma RIIa type R, in which the residue at position 131 Fc-gamma RIIa is Arg, several hundred times relative to the level of the same binding of Fc-gamma RIIb, which means that the selectivity of Fc-gamma RIIb binding is not increased compared to Fc-gamma RIIa type R (see, for example, PTL 15).

It is believed that only exposure to increased binding to Fc-gamma RIIa, but not increased binding to Fc-gamma RIIb, should affect cells, such as platelets, that express Fc-gamma RIIa but do not express Fc-gamma RIIb (see, e.g. NPL 17). For example, in the group of patients who were administered bevacizumab, an anti-VEGF antibody, it is known to have an increased risk of thromboembolism (see, for example, NPL 54). In addition, similarly, thromboembolism was detected in clinical studies of developed anti-CD40 ligand antibodies, and the clinical trial was discontinued (see, for example, NPL 55). Both versions of these antibodies have been studied in more recent studies using animal models and it has been suggested that the administered antibodies caused aggregation by Fc-gamma RIIa binding on platelets and led to blood clot formation (see, for example, NPL 56 and NPL 57). In systemic lupus erythematosus, which is an autoimmune disease, platelets are activated by an Fc-gamma RIIa dependent mechanism, and platelet activation has been described to correlate with the severity of symptoms (see, for example, NPL 58). The introduction of an antibody with increased ability to bind to Fc-gamma RIIa to those patients who already had a high risk of developing thromboembolism, the risk of developing thromboembolism may increase, which is extremely dangerous.

In addition, antibodies with increased ability to bind to Fc-gamma RIIa have been described to enhance macrophage-mediated antibody-dependent cellular phagocytosis (ADCP) (see, for example, NPL 59). When antigens to be bound by antibodies are phagocytosed by macrophages, it is believed that the antibodies themselves are also simultaneously phagocytosed. When antibodies are administered as pharmaceutical agents, it is expected that peptide fragments from the administered antibodies are also likely to be presented as an antigen, increasing the risk of producing antibodies against therapeutic antibodies (anti-therapeutic antibodies). More specifically, increased ability to bind to Fc-gamma RIIa may increase the risk of developing antibodies against therapeutic antibodies, and this may significantly reduce their value as pharmaceutical agents. In addition, Fc-gamma RIIb on dendritic cells has been suggested to be involved in peripheral tolerance by inhibiting dendritic cell activation induced by immune complexes formed between antigens and antibodies, or by suppressing antigen presentation to T cells via activation of Fc-gamma receptors (see ., for example, NPL 60). Since Fc-gamma RIIa is also expressed on dendritic cells, when antibodies that have an Fc with increased selectivity to bind to Fc-gamma RIIb were used as pharmaceutical agents, antigens could not be easily presented by dendritic cells and the like. due to increased selectivity to bind to Fc-gamma RIIb, and the risk of developing anti-drug antibodies could be relatively reduced. These antibodies may also be valuable in this regard.

More specifically, value as pharmaceuticals can be substantially reduced with increased ability to bind to Fc-gamma RIIa, resulting in an increased risk of thrombus formation from platelet aggregation and an increased risk of anti-therapeutic antibody production due to increased immunogenicity.

From this point of view, the above Fc variant with increased ability to bind to Fc-gamma RIIb is characterized by a significantly increased binding of R-type Fc-gamma RIIa compared to naturally occurring IgG1. Thus, its value as a pharmaceutical for patients who carry R-type Fc-gamma RIIa is greatly reduced. Types H and R Fc-gamma RIIa occur in Caucasians and African Americans with approximately the same frequency (see, for example, NPL 61 and NPL 62). Thus, when this Fc variant is used for the treatment of autoimmune diseases, the number of patients for whom it can be safely used to achieve its effectiveness as a pharmaceutical agent may be limited.

In addition, maturation of dendritic cells has been found to occur if the dendritic cells are deficient in Fc-gamma RIIb or if the interaction between Fc-gamma RIIb and the Fc region of an antibody to Fc-gamma RIIb is inhibited in dendritic cells (see, e.g., NPL 63 and NPL 64). In this study, it is suggested that RIIb Fc-gamma actively inhibits the maturation of dendritic cells in a stable state in the absence of inflammation or the like. and in the absence of activation. Fc-gamma RIIa is expressed on the surface of dendritic cells in addition to Fc-gamma RIIb; therefore, even if binding to inhibitory Fc-gamma RIIb is increased and if binding to activating Fc-gamma R such as Fc-gamma RIIa is also increased, maturation of dendritic cells may be enhanced as a result. More specifically, increasing not only Fc-gamma RIIb binding activity, but also the ratio of Fc-gamma RIIb-binding activity to Fc-gamma RIIa-binding activity is regarded as an important parameter in the development of immunosuppressive antibodies.

Thus, in developing a pharmaceutical that utilizes Fc-gamma RIIb binding-mediated immunosuppressive activity, there is a need for an Fc variant that not only has increased Fc-gamma RIIb binding activity, but also has the ability to bind to both Fc-gamma allotypes. RIIa H and R, which is maintained at a similar level or attenuated to a lower level compared to naturally occurring IgG1.

However, studies have now been published in which amino acid changes were introduced into the Fc region to increase the selectivity of binding to Fc-gamma RIIb (see, for example, NPL 65). However, all variants reported in this publication that had improved selectivity for Fc-gamma RIIb were found to have reduced ability to bind to Fc-gamma RIIb compared to naturally occurring IgG1. Thus, in fact, it seems difficult for these variants to induce an Fc-gamma RIIb mediated immunosuppressive response that is stronger than IgG1.

In addition, since the Fc-gamma of RIIb plays an important role for the above agonist antibodies, it is expected that an increase in their binding activity will lead to an increase in agonist activity. However, when binding to Fc-gamma RIIa is increased in a similar manner, activities such as ADCC activity and ADCP activity may be inadvertently displayed, and this may lead to side effects. From this point of view, it is preferable to be able to selectively increase the Fc-gamma RIIb-binding activity.

From these results it follows that in the production of therapeutic antibodies intended for the treatment of autoimmune diseases and cancer, in which Fc-gamma RIIb is involved, it is important that, compared to naturally occurring IgG1, the binding activity to both Fc-gamma RIIa allotypes is maintained. or decreased, and binding to Fc-gamma RIIb increased. However, Fc-gamma RIIb shares 93% extracellular sequence identity with Fc-gamma RIIa, which is one of the activating Fc-gamma Rs, and is structurally very similar. Fc-gamma RIIa, H-type and R-type allotypes are known, in which the amino acid at position 131 is His (H-type) or Arg (R-type), and each of them interacts differently with antibodies ( see, for example, NPL 66). Thus, it can be a difficult problem to generate an Fc region variant with increased selective binding to Fc-gamma RIIb over each of the Fc-gamma RIIa allotypes, including the ability to distinguish between highly homologous sequences in Fc-gamma RIIa and Fc-gamma RIIb. In light of these difficulties, several Fc variants have been identified to date that have selective binding activity for Fc-gamma RIIb over Fc-gamma RIIa by performing a wide-ranging analysis of amino acid modifications in the Fc region (see, e.g., PTL 16 , PTL 17, PTL 18, PTL 19 and PTL 20).

To date, a variant Fc region has been described with selective binding to Fc-gamma RIIb among human Fc-gamma R, but there is no data on a variant Fc region with selective binding to Fc-gamma RIIb among monkey Fc-gamma R. Given the absence of this Fc variant, the effects of the Fc variant selectively binding to Fc-gamma RIIb have not yet been extensively tested in monkeys.

In addition to the above, it is described that by modifying the charge of the amino acid residues that can be exposed on the surface of the antibody, resulting in an increase or decrease in the isoelectric point (pI) of the antibody, it is possible to regulate the half-life of the antibody in the blood (see, for example, PTL 21 and PTL 22) . These patents demonstrate that it is possible to prolong the plasma half-life of an antibody by lowering the pI of the antibody and vice versa.

In addition, it has been described that incorporation of an antigen into cells can be increased by modifying the charge of specific amino acid residues, primarily in the CH3 domain of an antibody, to increase its pI (see, for example, PTL 23). In addition, it is described that by modifying the charge of amino acid residues in the constant region (mainly in the CH1 domain) in order to reduce pI, the plasma half-life of the antibody can be prolonged (see, for example, PTL 24).

Link List

Patent Literature

PTL 1: US Patent No. 5827733

PTL 2: WO 1994/021681

PTL 3: US Patent No. 6096506

PTL 4: US Patent No. 7261893

PTL 5: US Patent No. 7320789

PTL 6: US Patent No. 7807159

PTL 7: US Patent No. 7888486

PTL 8: W02005/094446

PTL9: W02007/047112

PTL 10: W02010/070094

PTL 11: W02000/042072

PTL 12: W02006/019447

PTL 13: WO 2004/099249

PTL 14: W02004/029207

PTL 15: U.S. Patent Application Publication No. US 2009/0136485

PTL 16: W02012/115241

PTL 17: W02013/047752

PTL 18: WO 2013/125667

PTL 19: W02014/030728

PTL 20: WO 2014/163101

PTL 21: W02007/114319

PTL22: W02009/041643

PTL 23: W02014/145159

PTL 24: W02012/016227

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Brief summary of the invention

The technical problem underlying the present invention

The objective of the present invention was to develop antibodies to myostatin, polypeptides containing variants of the Fc regions, and methods of their use.

The solution of the problem

The present invention provides anti-myostatin antibodies and methods for their use. The invention also provides proteins containing a variant Fc region and methods of using the same.

In some embodiments, an isolated anti-myostatin antibody of the present invention binds to latent myostatin. In other embodiments, the antibody binds to an epitope within a fragment consisting of amino acids 21-100 of the myostatin propeptide (SEQ ID NO: 78). In some embodiments, the isolated anti-myostatin antibody of the present invention inhibits myostatin activation. In other embodiments, the antibody blocks the release of mature myostatin from latent myostatin. In other embodiments, the antibody blocks the proteolytic release of mature myostatin. In other embodiments, the antibody blocks the spontaneous release of mature myostatin. In other embodiments, the antibody does not bind to mature myostatin. In other embodiments, the antibody binds to the same epitope as the antibody shown in Table 13. In other embodiments, the antibody binds to the same epitope as the antibody that contains the VH and VL pair shown in Table 13. B in other embodiments, the antibody binds to the same epitope as the antibody shown in Table 2a. In other embodiments, the antibody binds to the same epitope as an antibody that contains the VH and VL pair shown in Table 2a. In other embodiments, the antibody binds to the same epitope as the antibody shown in Table 11a. In other embodiments, the antibody binds to the same epitope as an antibody that contains the VH and VL pair shown in Table 11a. In other embodiments, the antibody binds to the same epitope as the antibody shown in Table 2a, 11a, or 13. In other embodiments, the antibody binds to the same epitope as an antibody that contains the VH and VL pair shown in table 2a, 11a or 13.

In some embodiments, an isolated anti-myostatin antibody of the present invention binds latent myostatin with higher affinity at neutral pH than at acidic pH. In some embodiments, the anti-myostatin antibody binds latent myostatin with higher affinity at pH 7.4 than at pH 5.8. In some embodiments, an isolated anti-myostatin antibody of the present invention binds to a polypeptide fragment consisting of amino acids 21-100 of the myostatin propeptide (SEQ ID NO: 78) with higher affinity at pH 7.4 than at pH 5. ,8. In some embodiments, the antibody binds to the same myostatin epitope as the antibody shown in Table 13 with higher affinity at neutral pH than at acidic pH. In additional embodiments, the anti-myostatin antibody binds to the same epitope as the antibody shown in Table 13 with higher affinity at pH 7.4 than at pH 5.8. In other embodiments, the antibody binds to the same epitope as an antibody that contains the VH and VL pair shown in Table 13 with higher affinity at pH 7.4 than at pH 5.8. In some embodiments, the antibody binds to the same myostatin epitope as the antibody shown in Table 2a with higher affinity at neutral pH than at acidic pH. In some embodiments, the antibody binds to the same myostatin epitope as the antibody shown in Table 2a with higher affinity at pH 7.4 than at pH 5.8. In other embodiments, the antibody binds to the same epitope as an antibody that contains the VH and VL pair shown in Table 2a with higher affinity at pH 7.4 than at pH 5.8. In additional embodiments, the anti-myostatin antibody binds to the same epitope as the antibody shown in Table 11a with higher affinity at neutral pH than at acidic pH. In other embodiments, the antibody binds to the same myostatin epitope as the antibody shown in Table 11a at pH 7.4 than at pH 5.8. In other embodiments, the antibody binds to the same epitope as an antibody that contains the VH and VL pair shown in Table 11a with higher affinity at pH 7.4 than at pH 5.8. In additional embodiments, the anti-myostatin antibody binds to the same epitope as the antibody shown in Table 2a, 11a or 13 with higher affinity at neutral pH than at acidic pH. In other embodiments, the antibody binds to the same myostatin epitope as the antibody shown in Table 2a, 11a or 13 with higher affinity at pH 7.4 than at pH 5.8. In other embodiments, the antibody binds to the same epitope as an antibody that contains the VH and VL pair shown in Table 2a, 11a, or 13 with higher affinity at pH 7.4 than at pH 5.8.

In some embodiments, an isolated anti-myostatin antibody of the present invention competes for binding to latent myostatin with an antibody as defined herein. In some embodiments, an isolated anti-myostatin antibody of the present invention competes for binding to latent myostatin with an antibody shown in Table 13. In some embodiments, an isolated anti-myostatin antibody of the present invention competes for binding to latent myostatin , with an antibody that contains the VH and VL pair shown in Table 13. In some embodiments, the antibody competes for binding to latent myostatin with the antibody shown in Table 2a. In some embodiments, an isolated anti-myostatin antibody of the present invention competes for binding to latent myostatin with an antibody that contains the VH and VL pair shown in Table 2a. In some embodiments, the antibody competes for binding to latent myostatin with the antibody shown in Table 11a. In some embodiments, an isolated anti-myostatin antibody of the present invention competes for binding to latent myostatin with an antibody that contains the VH and VL pair shown in Table 11a. In additional embodiments, the anti-myostatin antibody competes for binding to latent myostatin with the antibody shown in Table 2a, 11a, or 13. In additional embodiments, the anti-myostatin antibody competes for binding to latent myostatin with an antibody that contains a pair of VH and VL, shown in Table 2a, 11a, or 13. In other embodiments, the anti-myostatin antibody binds latent myostatin with higher affinity at neutral pH than at acidic pH. In other embodiments, the anti-myostatin antibody binds latent myostatin with higher affinity at pH 7.4 than at pH 5.8. In other embodiments, the anti-myostatin antibody binds to a polypeptide fragment consisting of amino acids 21-100 of the myostatin propeptide (SEQ ID NO: 78) with higher affinity at pH 7.4 than at pH 5.8. Methods for assessing the ability of an antibody to compete with a reference antibody for binding to latent myostatin are described herein and are known in the art.

In some embodiments, the isolated anti-myostatin antibody of the present invention is a monoclonal antibody. In some embodiments, the isolated anti-myostatin antibody of the present invention is a human, humanized, or chimeric antibody. In some embodiments, the isolated anti-myostatin antibody of the present invention is an antibody fragment that binds to myostatin. In some embodiments, the isolated anti-myostatin antibody of the present invention is an antibody fragment that binds to latent myostatin. In some embodiments, the isolated anti-myostatin antibody of the present invention is an antibody fragment that binds to a polypeptide fragment consisting of amino acids 21-100 of the myostatin propeptide (SEQ ID NO: 78). In some embodiments, the isolated anti-myostatin antibody of the present invention is a full-length IgG-type antibody.

In some embodiments, an anti-myostatin antibody of the invention comprises:

(a) (I) HVR-H3 containing the amino acid sequence GVPAX ₁ SX ₂ GGDX ₃ in which X ₁ is Y or H, X ₂ is T or H, X ₃ is L or K (SEQ ID NO: 128), (II) HVR-L3 containing the amino acid sequence AGGYGGGX ₁ YA, wherein X ₁ is L or R (SEQ ID NO: 131), and (III) HVR-H2 containing the amino acid sequence IISX ₁ AGX ₂ X ₃ YX ₄ X ₅ X ₆ WAKX ₇ wherein X ₁ is Y or H, X ₂ is S or K, X ₃ is T, M or K, X ₄ is Y or K, X ₅ is A, M or E, X ₆ is S or E, X ₇ is G or K (SEQ IDNO: 127);

(b) (I) HVR-H1 containing the amino acid sequence X ₁ X ₂ DIS, in which X ₁ is S or H, X ₂ is Y, T, D or E (SEQ ID NO: 126), (II) HVR -H2 containing the amino acid sequence IISX ₁ AGX ₂ X ₃ YX ₄ X ₅ X ₆ WAKX ₇ in which X ₁ is Y or H, X 2 is S or K, _{X 3 is} T, M or K, X ₄ is Y or K, X ₅ is A, M or E, X ₆ is S or E, X ₇ is G or K (SEQ ID NO: 127), and (III) HVR-H3 containing the amino acid sequence GVPAX ₁ SX ₂ GGDX ₃ where X ₁ is Y or H, X ₂ is T or H, X ₃ is L or K (SEQ ID NO: 128);

(c) (I) HVR-H1 containing the amino acid sequence X ₁ X ₂ DIS, in which X ₁ is S or H, X ₂ is Y, T, D or E (SEQ ID NO: 126), (II) HVR -H2 containing the amino acid sequence IISX ₁ AGX ₂ X ₃ YX ₄ X ₅ X ₆ WAKX ₇ in which X ₁ is Y or H, X 2 is S or K, _{X 3 is} T, M or K, X ₄ is Y or K, X ₅ is A, M or E, X ₆ is S or E, X ₇ is G or K (SEQ ID NO: 127), (III) HVR-H3 containing the amino acid sequence GVPAX ₁ SX ₂ GGDX ₃ , in which X ₁ is Y or H, X ₂ is T or H, X ₃ is L or K (SEQ ID NO: 128), (IV) HVR-L1 containing the amino acid sequence X ₁ X ₂ SQX ₃ VX ₄ X ₅ X ₆ NWLS, wherein X ₁ is Q or T, X ₂ is S or T, X ₃ is S or E, X ₄ is Y or F, X ₅ is D or H, X ₆ is N, D, A or E (SEQ ID NO: 129); (V) HVR-L2 containing the amino acid sequence WAX ₁ TLAX ₂ in which X ₁ denotes S or E, X ₂ denotes S, Y, F or W (SEQ ID NO: 130); and (VI) HVR-L3 containing the amino acid sequence AGGYGGGX ₁ YA, in which X ₁ denotes L or R (SEQ ID NO: 131);

(d) (I) HVR-L1 containing the amino acid sequence X ₁ X ₂ SQX ₃ VX ₄ X ₅ X ₆ NWLS, in which X ₁ is Q or T, X ₂ is S or T, X ₃ is S or E, X ₄ is Y or F, X ₅ is D or H, X ₆ is N, D, A or E (SEQ ID NO: 129); (II) HVR-L2 containing the amino acid sequence WAX ₁ TLAX ₂ in which X ₁ denotes S or E, X ₂ denotes S, Y, F or W (SEQ ID NO: 130); and (III) HVR-L3 containing the amino acid sequence AGGYGGGX ₁ YA, wherein X ₁ is L or R (SEQ ID NO: 131). In some embodiments, the antibody specified in subparagraph (b) further comprises a FR1 heavy chain variable domain framework region comprising one of the amino acid sequences of SEQ ID NOs: 132-134; FR2 containing one of the amino acid sequences of SEQ ID NO: 135-136; FR3 containing the amino acid sequence of SEQ ID NO: 137; and FR4 comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the antibody of subparagraph (d) further comprises an FR1 light chain variable domain framework region comprising the amino acid sequence of SEQ ID NO: 139; FR2 containing one of the amino acid sequences of SEQ ID NO: 140-141; FR3 containing one of the amino acid sequences of SEQ ID NO: 142-143; and FR4 containing the amino acid sequence of SEQ ID NO: 144.

In some embodiments, an isolated anti-myostatin antibody of the present invention comprises (a) an HVR-H3 comprising the amino acid sequence GVPAX ₁ SX ₂ GGDX ₃ , wherein X ₁ is Y or H, X ₂ is T or H, X ₃ is L or K (SEQ ID NO: 128), (b) HVR-L3 containing the amino acid sequence AGGYGGGX ₁ YA, wherein X ₁ is L or R (SEQ ID NO: 131), and (c) HVR-H2 containing the amino acid sequence IISX ₁ AGX ₂ X ₃ YX ₄ X ₅ X ₆ WAKX ₇ in which X ₁ is Y or H, X ₂ is S or K, X ₃ is T, M or K, X ₄ is Y or K , X ₅ is A, M or E, X ₆ is S or E, X ₇ is G or K (SEQ ID NO: 127).

In some embodiments, the isolated anti-myostatin antibody of the present invention comprises (a) an HVR-H1 comprising the amino acid sequence X ₁ X ₂ DIS, wherein X ₁ is S or H, X ₂ is Y, T, D, or E (SEQ ID NO: 126), (b) HVR-H2 containing the amino acid sequence IISX ₁ AGX ₂ X ₃ YX ₄ X ₅ X ₆ WAKX ₇ in which X ₁ is Y or H, X ₂ is S or K, X ₃ is T, M or K, X ₄ is Y or K, X ₅ is A, M or E, X ₆ is S or E, X ₇ is G or K (SEQ ID NO: 127), and (c) HVR-H3 containing the amino acid sequence GVPAX ₁ SX ₂ GGDX ₃ in which X ₁ is Y or H, X ₂ is T or H, X ₃ is L or K (SEQ ID NO: 128). In other embodiments, the antibody comprises a FR1 heavy chain variable domain framework region comprising one of the amino acid sequences of SEQ ID NOs: 132-134; FR2 containing one of the amino acid sequences of SEQ ID NO: 135-136; FR3 containing the amino acid sequence of SEQ ID NO: 137; and FR4 comprising the amino acid sequence of SEQ ID NO: 138. In other embodiments, the antibody further comprises (a) HVR-L1 comprising the amino acid sequence X ₁ X ₂ SQX ₃ VX ₄ X ₅ X ₆ NWLS, wherein X ₁ is Q or T, X ₂ is S or T, X ₃ is S or E, X ₄ is Y or F, X ₅ is D or H, X ₆ is N, D, A or E (SEQ ID NO: 129); (b) HVR-L2 containing the amino acid sequence WAX ₁ TLAX ₂ in which X ₁ denotes S or E, X ₂ denotes S, Y, F or W (SEQ ID NO: 130); and (c) HVR-L3 containing the amino acid sequence AGGYGGGX ₁ YA, wherein X ₁ is L or R (SEQ ID NO: 131).

In some embodiments, the isolated anti-myostatin antibody of the present invention comprises (a) an HVR-L1 comprising the amino acid sequence X ₁ X ₂ SQX ₃ VX ₄ X ₅ X ₆ NWLS, wherein X ₁ is Q or T, X ₂ is S or T, X ₃ is S or E, X ₄ is Y or F, X ₅ is D or H, X ₆ is N, D, A or E (SEQ ID NO: 129); (b) HVR-L2 containing the amino acid sequence WAX ₁ TLAX ₂ in which X ₁ denotes S or E, X ₂ denotes S, Y, F or W (SEQ ID NO: 130); and (c) HVR-L3 containing the amino acid sequence AGGYGGGX ₁ YA, wherein X ₁ is L or R (SEQ ID NO: 131). In a further embodiment, the antibody further comprises an FR1 light chain variable domain framework region comprising the amino acid sequence of SEQ ID NO: 139; FR2 containing one of the amino acid sequences of SEQ ID NO: 140-141; FR3 containing one of the amino acid sequences of SEQ ID NO: 142-143; and FR4 containing the amino acid sequence of SEQ ID NO: 144.

In some embodiments, the isolated anti-myostatin antibody of the present invention comprises an FR1 heavy chain variable domain framework region comprising one of the amino acid sequences of SEQ ID NOs: 132-134; FR2 containing one of the amino acid sequences of SEQ ID NO: 135-136; FR3 containing the amino acid sequence of SEQ ID NO: 137; and FR4 comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the isolated anti-myostatin antibody of the present invention comprises an FR1 light chain variable domain framework region comprising the amino acid sequence of SEQ ID NO: 139; FR2 containing one of the amino acid sequences of SEQ ID NO: 140-141; FR3 containing one of the amino acid sequences of SEQ ID NO: 142-143; and FR4 containing the amino acid sequence of SEQ ID NO: 144.

In some embodiments, an isolated anti-myostatin antibody of the present invention comprises (a) a VH sequence that is at least 95% identical to one of the amino acid sequences of SEQ ID NOs: 13, 16-30, 32-34, and 86-95 ; (b) a VL sequence at least 95% identical to one of the amino acid sequences of SEQ ID NOs: 15, 31, 35-38 and 96-99; or (c) a VH sequence as specified in subparagraph (a) and a VL sequence as specified in subparagraph (b). In other embodiments, the antibody comprises a VH sequence as set forth in one of SEQ ID NOs: 13, 16-30, 32-34, and 86-95. In other embodiments, the antibody comprises a VL sequence as set forth in one of SEQ ID NOs: 15, 31, 35-38, and 96-99. In some embodiments, the antibody comprises a VH sequence as set forth in one of SEQ ID NOs: 13, 16-30, 32-34, and 86-95. In other embodiments, the antibody comprises the VH sequence shown in one of SEQ ID NOs: 13, 16-30, 32-34, and 86-95; and the VL sequence shown in one of SEQ ID NOs: 15, 31, 35-38 and 96-99.

The invention also relates to isolated nucleic acids encoding an anti-myostatin antibody of the present invention. The invention also relates to host cells that contain the nucleic acid of the present invention. The invention also relates to a method for producing an antibody comprising culturing a host cell of the present invention so as to obtain an antibody.

In some aspects of the invention, a method for producing an antibody to myostatin is provided, comprising (a) culturing a host cell of the present invention so as to obtain an antibody; or (b) immunizing the animal with a polypeptide, wherein the polypeptide comprises a region corresponding to amino acids at positions 21-100 of the myostatin propeptide (SEQ ID NO: 78).

The invention also provides a method for producing antibodies to myostatin. In some embodiments, the method comprises immunizing an animal with a polypeptide, wherein the polypeptide comprises a region corresponding to amino acids at positions 21-100 of the myostatin propeptide (SEQ ID NO: 78).

The invention also relates to a pharmaceutical composition which contains an anti-myostatin antibody of the present invention and a pharmaceutically acceptable carrier.

The anti-myostatin antibodies of the present invention can be used as a medicine. In some embodiments, the antibody is used to prepare a medicament for: (a) treating a muscle wasting syndrome (disease); (b) increase in muscle tissue mass; (c) increasing the strength of muscle tissue; or (d) reducing the accumulation of body fat. In some embodiments, the anti-myostatin antibodies of the present invention can be used to treat muscle wasting syndrome. The anti-myostatin antibodies of the present invention can be used to increase muscle mass. The anti-myostatin antibodies of the present invention can be used to increase the strength of muscle tissue. The anti-myostatin antibodies of the present invention can be used to reduce the accumulation of body fat.

In some embodiments, an anti-myostatin antibody provided herein is for use in: (a) treating muscle wasting syndrome; (b) increase in muscle tissue mass; (c) increasing the strength of muscle tissue; or (d) reducing the accumulation of body fat.

The anti-myostatin antibodies of the present invention can be used in the preparation of a medicament. In some embodiments, the antibody is used to prepare a medicament for the treatment of: (a) muscle wasting syndrome; (b) increase in muscle tissue mass; (c) increasing the strength of muscle tissue; or (d) reducing the accumulation of body fat. In some embodiments, the drug is for the treatment of muscle wasting syndrome. In some embodiments of the invention, the drug is intended to increase the mass of muscle tissue. In some embodiments of the invention, the drug is intended to increase the strength of muscle tissue. In some embodiments of the invention, the drug is intended to reduce the accumulation of body fat.

The invention also provides a method of treating an individual having a muscle wasting syndrome. In some embodiments, the method includes administering to the subject an effective amount of an anti-myostatin antibody of the present invention. The invention also provides a method for increasing the mass of muscle tissue in an individual. In some embodiments, the method includes administering to an individual an effective amount of an anti-myostatin antibody of the present invention to increase muscle mass. The invention also provides a method for increasing the strength of muscle tissue in an individual. In some embodiments, the method includes administering to an individual an effective amount of an anti-myostatin antibody of the present invention to increase muscle strength. The invention also provides a method for reducing the accumulation of body fat in an individual. In some embodiments, the method includes administering to an individual an effective amount of an anti-myostatin antibody of the present invention to reduce body fat accumulation.

The invention also relates to polypeptides containing variants of the Fc regions, and methods for their preparation and use.

One of the embodiments of the invention is Fc-gamma RIIb-binding polypeptides containing variants of the Fc regions, and methods of their use. In some embodiments, a variant Fc region with increased Fc-gamma RIIb binding activity of the present invention contains at least one amino acid change in the parent Fc region. In other embodiments, the ratio [KD value of the parent Fc region for simian Fc-gamma R11b]/[KD value of the variant Fc region for simian Fc-gamma RIIb] is 2.0 or greater. In other embodiments, the ratio [KD value of parental Fc region for simian Fc-gamma RIIIa]/[KD value of variant Fc region for simian Fc-gamma RIIIa] is 0.5 or less. In other embodiments, the ratio [KD value of the parental Fc region for human Fc-gamma RIIb]/[KD value of the variant Fc region for human Fc-gamma RIIb] is 2.0 or greater. In other embodiments, the ratio [KD value of the parental Fc region for human Fc-gamma RIIIa]/[KD value of the variant Fc region for human Fc-gamma RIIIa] is 0.5 or less. In other embodiments, the ratio [KD value of parental Fc region for human Fc-gamma RIIa (H-type)]/[KD value of variant Fc region for human Fc-gamma RIIa (H-type)] is 5.0 or less. In other embodiments, the ratio [KD value of parent Fc region for human Fc gamma RIIa (R) type]/[KD value of variant Fc region for human Fc gamma RIIa (R type)] is 5.0 or less. In another embodiment of the invention, the KD value of the Fc region variant for the simian Fc-gamma RIIb is 1.0 x 10 ^-6 M or less. In another embodiment of the invention, the KD value of the Fc region variant for the simian Fc-gamma RIIIa is 5.0×10 ^-7 M or more. In another embodiment, the KD value of the Fc region variant for human Fc-gamma RIIb is 2.0 x 10 ^-6 M or less. In another embodiment, the KD value of the Fc region variant for human Fc-gamma RIIIa is 1.0 x 10 ^-6 M or greater. In another embodiment of the invention, the KD value of the Fc region variant for human Fc-gamma RIIa (H-type) is 1.0×10 M ^-7 or more. In another embodiment of the invention, the KD value of the Fc region variant for human Fc-gamma RIIa (R-type) is 2.0×10 ^-7 M or more.

In some embodiments, a variant Fc region with enhanced Fc-gamma RIIb binding activity of the present invention contains at least one amino acid change at at least one position selected from the group consisting of: 231, 232, 233 , 234, 235, 236, 237, 238, 239, 264, 266, 267, 268, 271, 295, 298, 325, 326, 327, 328, 330, 331, 332, 334 and 396, according to EU numbering.

In other embodiments, the Fc region variant with increased Fc-gamma RIIb binding activity comprises at least two amino acid changes, including: (a) one amino acid change at position 236, and (b) at least one amino acid change of at least at one position selected from the group consisting of: (I) positions: 231, 232, 233, 234, 235, 237, 238, 239, 264, 266, 267, 268, 271, 295, 298, 325, 326, 327 , 328, 330, 331, 332, 334 and 396; (II) provisions: 231, 232, 235, 239, 268, 295, 298, 326, 330 and 396; or (III) provisions 268, 295, 326 and 330; according to EU numbering.

In other embodiments, the Fc region variant with increased Fc-gamma RIIb binding activity comprises at least two amino acid changes, including: (a) one amino acid change at position 236, and (b) at least one amino acid change of at least at one position selected from the group consisting of: 231, 232, 233, 234, 235, 237, 238, 239, 264, 266, 267, 268, 271, 295, 298, 325, 326, 327, 328, 330 , 331, 332, 334 and 396, according to the EU numbering.

In other embodiments, the Fc region variant with increased Fc-gamma RIIb binding activity comprises at least two amino acid changes, including: (a) one amino acid change at position 236, and (b) at least one amino acid change of at least in one position selected from the group consisting of: 231, 232, 235, 239, 268, 295, 298, 326, 330 and 396, according to EU numbering.

In other embodiments, the Fc region variant with increased Fc-gamma RIIb binding activity comprises at least two amino acid changes, including: (a) one amino acid change at position 236, and (b) at least one amino acid change of at least in one position selected from the group consisting of: 268, 295, 326 and 330, according to EU numbering.

In some embodiments, the Fc region variant with increased Fc-gamma RIIb binding activity comprises at least one amino acid selected from the group consisting of: (a) Asp, Glu, Phe, Gly, His, Ile, Lys, Leu , Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, Tyr at position 231; (b) Ala, Asp, Glu, Phe, Gly, His, He, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, Tyr at position 232; (c) Asp at position 233; (d) Trp, Tyr at position 234; (e) Trp at position 235; (f) Ala, Asp, Glu, His, Ile, Leu, Met, Asn, Gln, Ser, Thr, Val at position 236; (g) Asp, Tyr at position 237; (h) Glu, Ile, Met, Gln, Tyr at position 238; (i) Ile, Leu, Asn, Pro, Val at position 239; (j) Ile at position 264; (k) Phe at position 266; (m) Ala, His, Leu at position 267; (m) Asp, Glu at position 268; (o) Asp, Glu, Gly at position 271; (p) Leu at position 295; (p) Leu at position 298; (c) Glu, Phe, Ile, Leu at position 325; (r) Thr at position 326; (y) Ile, Asn at position 327; (f) Thr at position 328; (x) Lys, Arg at position 330; (c) Glu at position 331; (h) Asp at position 332; (w) Asp, Ile, Met, Val, Tyr at position 334; and (y) Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gin, Arg, Ser, Thr, Val, Trp, Tyr at position 396; according to EU numbering.

In other embodiments, the variant Fc region with increased Fc-gamma RIIb binding activity contains at least one amino acid selected from the group consisting of: (a) Gly, Thr at position 231; (b) Asp at position 232; (c) Trp at position 235; (d) Asn, Thr at position 236; (e) Val at position 239; (e) Asp, Glu at position 268; (g) Leu at position 295; (h) Leu at position 298; (i) Thr at position 326; (j) Lys, Arg at position 330 and (k) Lys, Met at position 396; according to EU numbering.

Another embodiment of the invention is a polypeptide that contains a variant Fc region with an increased isoelectric point (pI) and a method of using it. In some embodiments, a polypeptide that contains an elevated pI variant Fc region includes at least two amino acid changes in the parent Fc region. In other embodiments, each of the amino acid changes increases the isoelectric point (pI) of the variant Fc region relative to the parent Fc region. In other embodiments of the invention, the amino acid may be exposed on the surface of the variant Fc region. In other embodiments, the polypeptide comprises a variant Fc region and an antigen binding domain. In other embodiments of the invention, the antigen-binding activity of the antigen-binding domain varies depending on the concentration of ions. In other embodiments, the increased pI variant Fc region of the present invention contains at least two amino acid changes at at least two positions selected from the group consisting of: 285, 311, 312, 315, 318, 333 , 335, 337, 341, 342, 343, 384, 385, 388, 390, 399, 400, 401, 402, 413, 420, 422 and 431, according to EU numbering. In other embodiments, the elevated pI variant Fc region contains Arg or Lys at each of the selected positions.

In some embodiments, the Fc region variant of the present invention contains the amino acid changes shown in Tables 14-30.

In some embodiments, the polypeptide comprises an Fc region variant of the present invention. In other embodiments, the parental Fc region is derived from human IgG1. In other embodiments, the polypeptide is an antibody. In other embodiments, the polypeptide is an Fc-containing fusion protein.

The invention provides a polypeptide containing one of the amino acid sequences of SEQ ID NO: 229-381.

The invention also relates to an isolated nucleic acid which encodes a polypeptide containing a variant Fc region according to the invention. The invention also relates to a host cell that contains the nucleic acid of the present invention. The invention also relates to a method for producing a polypeptide that contains a variant Fc region, including the cultivation of the host proposed in the present invention, so as to obtain a polypeptide.

The invention also relates to a pharmaceutical composition that contains a polypeptide containing a variant of the Fc region proposed in the present invention, and a pharmaceutically acceptable carrier.

Brief description of the drawings

The drawings show:

in fig. 1 Inhibition of proteolytic activation of latent myostatin by anti-latent myostatin antibody as illustrated in Example 3. The activity of active myostatin released from latent myostatin by the action of BMP1 protease was measured in the presence of anti-latent myostatin antibody using the HEK Blue assay;

in fig. 2 Inhibition of spontaneous activation of latent myostatin by anti-latent myostatin antibody as illustrated in Example 4. The activity of active myostatin released from latent myostatin by incubation at 37° C. was measured in the presence of anti-latent myostatin antibody using the HEK Blue assay;

in fig. 3 - binding of antibodies to latent myostatin with the propeptide domain, as illustrated in example 5;

in fig. 4 shows the results of a Western blot analysis against myostatin propeptide as illustrated in Example 6. Proteolytic cleavage of myostatin propeptide by BMP1 was assessed in the presence and absence of anti-latent myostatin antibody;

in fig. 5A-5C are BIACORE®-derived sensograms characterizing the binding of anti-latent myostatin antibody MST1032-G1m to human latent myostatin (A), monkey cynomolgus latent myostatin (B), and mouse latent myostatin (C), as illustrated in Example 7;

in fig. 6A shows the in vivo efficacy of anti-latent myostatin antibody and anti-mature myostatin antibody on muscle mass and fat mass as illustrated in Example 8. ) was administered to SCID mice and total lean (fat-free) body mass (or lean body mass: LBM) was assessed;

in fig. 6B In vivo efficacy of anti-latent myostatin antibody and anti-mature myostatin antibody on muscle mass and fat mass as illustrated in Example 8. ) was administered to SCID mice and the change in total body fat mass from day 0 to day 14 was assessed;

in fig. 6B shows the in vivo efficacy of anti-latent myostatin antibody and anti-mature myostatin antibody on muscle mass and fat mass as illustrated in Example 8. ) was administered to SCID mice and calf and quadriceps muscle mass was assessed;

in fig. 7A Comparison of the in vivo efficacy of several anti-myostatin antibodies as illustrated in Example 9. An anti-latent myostatin antibody (MST1032-G1m; designated as MST1032 in the drawings) or an anti-mature myostatin antibody (41C1E4, REGN, OGD or MYO-029) was injected with SCID mice and assessed the total lean body mass;

in fig. 7B Comparison of the in vivo efficacy of several anti-myostatin antibodies as illustrated in Example 9. An anti-latent myostatin antibody (MST1032-G1m; designated as MST1032 in the drawings) or an anti-mature myostatin antibody (41C1E4, REGN, OGD or MYO-029) was injected with SCID mice and evaluated the force of compression;

in fig. 7B Comparison of the in vivo efficacy of several anti-myostatin antibodies as illustrated in Example 9. An anti-latent myostatin antibody (MST1032-G1m; designated as MST1032 in the drawings) or an anti-mature myostatin antibody (41C1E4, REGN, OGD or MYO-029) was injected with SCID mice and assessed the change in total body fat from day 0 to day 14;

in fig. 8 - inhibition of proteolytic and spontaneous activation of latent myostatin by a humanized antibody to latent myostatin, as illustrated in example 10. The activity of active myostatin released from latent myostatin by the action of BMP1 protease (proteolytic) or incubation at 37°C without BMP1 (spontaneous) was measured in the presence of anti-latent myostatin antibody using the HEK Blue assay;

in fig. 9 - BIACORE® generated sensorgrams characterizing the binding of variants to a substituted histidine of an anti-latent myostatin antibody as illustrated in Example 11. The antibody/antigen complexes were allowed to dissociate at pH 7.4 followed by further dissociation at pH 5.8 (indicated by an arrow) to study pH-dependent interactions. The following antibodies were tested in this experiment: Ab001 (curve indicated by black solid line), Ab002 (curve indicated by black dotted line), Ab003 (curve indicated by black dotted line), Ab004 (curve indicated by gray dashed line), Ab005 (curve indicated by gray dashed line). , indicated by a gray solid line), Ab006 (curve indicated by gray long strokes) and Ab007 (curve indicated by black long strokes);

Figure 10 - Inhibition of proteolytic and spontaneous activation of latent myostatin by pH-dependent antibodies to latent myostatin, as illustrated in example 13. Activity of active myostatin released from latent myostatin by the action of BMP1 protease (proteolytic) or incubation at 37°C without BMP1 (spontaneous ) was measured in the presence of anti-latent myostatin antibody using the HEK Blue assay. In the drawing, antibodies MS1032LO01-SG1, MS1032LO02-SG1, MS1032LO03-SG1 and MS1032LO04-SG1 are designated as MSLO-01, MSLO-02, MSLO-03 and MSLO-04, respectively. Inhibition of proteolytic and spontaneous activation of latent myostatin, comparable to MS1032LO00-SG1, was achieved using MS1032LO01-SG1, MS1032LO02-SG1, MS1032LO03-SG1 and MS1032LO04-SG1:

in fig. 11A-11E are BIACORE®-derived sensorgrams characterizing the binding of a pH-dependent antibody to latent myostatin as illustrated in Example 14. Kinetic parameters of MST1032-SG1 (A), MS1032LO00-SG1 (B), MS1032LO01-SG1 (C), MS1032LO02-SG1 (D), MS1032LO03-SG1 (D) and MS1032LO04-SG1 (E) were measured at neutral pH and acidic pH;

in fig. 12 is a time course of plasma myostatin concentration following intravenous administration of an anti-myostatin antibody to mice as illustrated in Example 15. The effects of Fc-gamma R-mediated cell uptake of antibody/antigen complexes on myostatin clearance in vivo were assessed by comparing anti-myostatin antibodies having the ability to bind with Fc-gamma R (MS1032LO00-SG1), and anti-myostatin antibodies with eliminated ability to bind to Fc-gamma R (MS1032LO00-F760);

in fig. 13 is a time course of plasma myostatin concentration following intravenous administration of anti-myostatin antibody to mice, as illustrated in Example 16. MS1032LO01-F760) and a pH independent anti-myostatin antibody (MS1032LO00-SG1 or MS1032LO00-F760);

in fig. 14A-14E are in vivo efficacy of pH-dependent and pH-independent latent myostatin antibodies as illustrated in Example 17. myostatin (MS1032LO00-SG1; designated as MSLO0 in the figure) was administered to SCID mice and total lean body mass (A), total body fat mass (B), quadriceps mass (C) were assessed; gastrocnemius muscle mass (G) and compression force (D);

in fig. 15: Binding activity of latent myostatin antibody MST1032 to latent myostatin and GDF11 as illustrated in Example 19;

in fig. 16 - Inhibitory activity of the latent myostatin antibody MST1032 against proteolytic and spontaneous activation of GDF11, as illustrated in example 20. The activity of active GDF11 released by the action of BMP1 protease (proteolytic) or incubation at 37°C without BMP1 (spontaneous) was measured in the presence of latent myostatin antibodies using the HEK Blue assay;

in fig. 17 Inhibition of proteolytic activation of latent myostatin by anti-latent myostatin antibody as illustrated in Example 22. The activity of active myostatin released by BMP1 protease was measured in the presence of anti-latent myostatin antibody using the HEK Blue assay;

in fig. 18 is a time course of plasma myostatin concentration following intravenous administration of anti-latent myostatin antibody to mice as illustrated in Example 23. Effects of pH dependence on myostatin clearance in vivo were assessed by comparing pH-independent anti-myostatin latent antibody (MS1032LO00-SG1) and various pH-dependent antibodies to latent myostatin (MS1032LO01-SG1, MS1032LO06-SG1, MS1032LO11-SG1, MS1032LO18-SG1, MS1032LO19-SG1, MS1032LO21-SG1 and MS1032LO25-SG1);

in fig. 19A and 19B are time-lapse plasma myostatin concentrations following intravenous administration of anti-latent myostatin antibody to cynomolgus monkeys as illustrated in Example 24. antibodies to latent myostatin (MS1032LO00-SG1) and pH-dependent antibodies to latent myostatin with engineered Fc (MS1032LO06-SG1012, MS1032LO06-SG1016, MS1032LO06-SG1029, MS1032LO06-SG1031, MS1032LO06-SG1030, MS10233). (B) The effect of Fc construction on myostatin clearance in vivo was assessed by comparing antibodies to latent myostatin (MS1032LO19-SG1079, MS1032LO19-SG1071, MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081 and MS1032LO19-SG1077);

in fig. 20A, as well as in FIG. 20B-20I In vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip strength and body fat mass as illustrated in Example 25. MS1032LO06-SG1, MS1032LO11-SG1 and MS1032LO18-SG1 were administered Scid mice and lean body mass was evaluated (A);

in fig. 20B, as well as in FIG. 20A, 20B-20I show in vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip force, and body fat mass as illustrated in Example 25. MS1032LO06-SG1, MS1032LO19-SG1, and MS1032LO25-SG1 was administered to Scid mice and lean body mass was assessed (B);

in fig. 20B, as well as in FIG. 20A-B, 20D-I show the in vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip strength and body fat mass as illustrated in Example 25. MS1032LO01-SG, MS1032LO06-SG1 and MS1032LO11 -SG1 was administered to Scid mice and lean body mass was assessed (B);

in fig. 20D, as well as in Fig. 20A-C, 20E-I - In vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip force, and body fat mass as illustrated in Example 25. MS1032LO06-SG1, MS1032LO11-SG1, and MS1032LO18 -SG1 was administered to Scid mice and the grip strength (G) was assessed;

in fig. 20D, as well as in Fig. 20A-D, 20E-20I show in vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip strength and body fat mass as illustrated in Example 25. MS1032LO06-SG1, MS1032LO19-SG1 and MS1032LO25 -SG1 was administered to Scid mice and the grip strength (D) was assessed;

in fig. 20E, as well as in FIG. 20A-E, 20G-I are in vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip force and body fat mass as illustrated in Example 25. MS1032LO01-SG, MS1032LO06-SG1 and MS1032LO11 -SG1 was administered to Scid mice and grip strength was evaluated (E);

in fig. 20G, as well as in FIG. 20A-E, 203-I are in vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip force and body fat mass as illustrated in Example 25. MS1032LO06-SG1, MS1032LO11-SG1 and MS1032LO18 -SG1 was administered to Scid mice and body fat mass (W) was assessed;

in fig. 203 and also in FIG. 20A-G, 20I - In vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip force and body fat mass as illustrated in Example 25. MS1032LO06-SG1, MS1032LO19-SG1 and MS1032LO25-SG1 administered to Scid mice and assessed for body fat (3);

in fig. 20I, as well as in FIG. 20A-3 In vivo efficacy of anti-latent myostatin antibodies (MS1032 variants) on lean body mass (LBM), grip strength and body fat mass as illustrated in Example 25. MS1032LO01-SG, MS1032LO06-SG1 and MS1032LO11-SG1 were treated with Scid mice and estimated body fat mass (I);

in fig. 21 Inhibitory activity against latent myostatin activation of antibodies to latent myostatin as illustrated in Example 26. Amounts of mature myostatin released from latent myostatin by BMP1 protease were measured in the presence of antibodies to latent myostatin (MST1032, MST1504, MST1538, MST1551, MST1558 , MST1572 and MST1573);

in fig. 22A is a schematic diagram of latent myostatin fragments of 100 amino acids each designed for epitope mapping of antibodies to latent myostatin as illustrated in Example 26;

in fig. 22B shows the results of a Western blot analysis against GST labeled human latent myostatin (GST-hMSTN) fragments using an anti-GST antibody as illustrated in Example 26. Each lane corresponds to: 1, GST-hMSTN 1-100 aa; 2, GST-hMSTN 21-120 ac; 3, GST-hMSTN 41-140 aa; 4, GST-hMSTN 61-160 ac; 5, GST-hMSTN 81-180 ac; 6, GST-hMSTN 101-200ac; 7, GST-hMSTN 121-220 ac; 8, GST-hMSTN 141-241 aa; 9, GST control;

in fig. 22B - Western blot analysis results against GST-labeled human latent myostatin (GST-hMSTN) fragments using antibodies to latent myostatin (MST1032, MST1538, MST1572 and MST1573) as illustrated in Example 26. Each lane corresponds to: 1, GST- hMSTN 1-100 ak; 2, GST-hMSTN 21-120 ac; 3, GST-hMSTN 41-140 aa; 4, GST-hMSTN 61-160 ac; 5, GST-hMSTN 81-180 ac; 6, GST-hMSTN 101-200ac; 7, GST-hMSTN 121-220 ac; 8, GST-hMSTN 141-241 aa; 9, GST control; 10, human latent myostatin (100 ng);

in fig. 22D is a summary of Western blot D analysis and predicted epitope positions for latent myostatin antibodies (MST1032, MST1538, MST1572 and MST1573) as illustrated in Example 26;

in fig. 23 - comparative analysis of the primary structure of the amino acid sequences of simian (cynomolgus, (cyno)) Fc-gamma RIIa 1, Fc-gamma RIIa2, Fc-gamma RIIa3, Fc-gamma RIIb, human Fc-gamma RIIaH, Fc-gamma RIIaR and Fc- gamma RIIb. The squared regions correspond to putative residues that interact with the Fc domain;

in fig. 24 is a time course of total plasma myostatin concentration following intravenous administration of an anti-myostatin antibody with an Fc variant with increased ability to bind Fc-gamma RIIb to all human Fc-gamma R transgenic mice as illustrated in Example 28. The effects of Fc variants with increased the ability to bind to Fc-gamma RIIb to eliminate the antigen by human Fc-gamma RIIb;

in fig. 25 is a time course of plasma antibody concentration following intravenous administration of an anti-myostatin antibody with an Fc variant with increased ability to bind to Fc-gamma RIIb to all human Fc-gamma R transgenic mice as illustrated in Example 28. The effects of Fc variants with increased ability were assessed. bind to Fc-gamma RIIb on antibody pharmacokinetics;

in fig. 26A and 26B are time-dependent plasma myostatin concentrations following intravenous administration of anti-latent myostatin antibodies to the Cynomolgus monkey as illustrated in Example 29. latent myostatin (MS1032LO00-SG1) and pH-dependent antibodies to latent myostatin with an engineered Fc region (MS1032LO06-SG1012, MS1032LO06-SG1016, MS1032LO06-SG1029, MS1032LO06-SG1031, MS1032LO06-SG10SG1033, MS10323). (B) The effect of Fc construction on myostatin clearance in vivo was assessed by comparing antibodies to latent myostatin (MS1032LO19-SG1079, MS1032LO19-SG1071, MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081, and MS1032LO19-SG1077);

in fig. 27A is a time course of total plasma myostatin concentration following intravenous administration of an anti-myostatin antibody with increased pI Fc variant to human FcRn transgenic mice as illustrated in Example 30. The effects of elevated pI Fc variants on antigen elimination were assessed;

in fig. 27B is a time course of plasma antibody concentration following intravenous administration of an anti-myostatin antibody with elevated pI Fc variant to human FcRn transgenic mice as illustrated in Example 30. The effects of elevated pI Fc variants on antibody pharmacokinetics were assessed;

in fig. 28A is time course of total plasma myostatin concentration following intravenous administration of an anti-myostatin antibody with elevated pI Fc variant to human FcRn transgenic mice as illustrated in Example 30. The effects of elevated pI Fc variants on antigen clearance were assessed. In this assay, human normal immunoglobulin was over-administered together with an anti-myostatin antibody to mimic the situation in human plasma;

in fig. 28B is a time course of plasma antibody concentration following intravenous administration of an anti-myostatin antibody with elevated pI Fc variant to human FcRn transgenic mice as illustrated in Example 30. The effects of elevated pI Fc variants on antibody pharmacokinetics were assessed. In this assay, human normal immunoglobulin was over-administered together with an anti-myostatin antibody to mimic the situation in human plasma;

in fig. 29 is a time course of total plasma myostatin concentration following intravenous administration of an anti-myostatin antibody with an Fc variant with increased ability to bind to Fc-gamma RIIb human Fc-gamma RIIb transgenic mice as illustrated in Example 31. The effects of Fc variants with increased ability were assessed. bind to Fc-gamma RIIb to eliminate the antigen by human Fc-gamma RIIb;

in fig. 30 is a time course of plasma antibody concentration following intravenous administration of an anti-myostatin antibody with enhanced Fc variant binding to Fc-gamma RIIb human Fc-gamma RIIb transgenic mice as illustrated in Example 31. The effects of Fc variants with enhanced binding were assessed. with Fc-gamma RIIb on antibody pharmacokinetics;

in fig. 31 - Cell imaging assay results of anti-myostatin antibodies with an Fc variant with increased ability to bind to Fc-gamma RIIb as illustrated in Example 33. Each antibody was complexed with fluorescently labeled myostatin and intracellular uptake of the antigen-antibody complex was measured in cells expressing human Fc-gamma RIIb.

Description of embodiments of the invention

The techniques and procedures described or referenced herein are generally well known to those skilled in the art and are widely used using conventional methodologies such as those described by Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 2001; Current Protocols in Molecular Biology (ed. R.M. Ausubel et al., 2003); Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (ed. M.J. MacPherson, B.D. Hames and G.R. Taylor, 1995), Antibodies, A Laboratory Manual, ed. Harlow and Lane, 1988 and Animal Cell Culture (ed. R.I. Freshney 1987); Oligonucleotide Synthesis (ed. M.J. Gait, 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (ed. J.E. Cellis, 1998) from Academic Press; Animal Cell Culture (ed. R.I. Freshney, 1987); Introduction to Cell and Tissue Culture (J. P. Mather and P. E. Roberts, 1998), Plenum Press; Cell and Tissue Culture: Laboratory Procedures (ed. A. Doyle, J. B. Griffiths and D. G. Newell, 1993-8), J. Wiley and Sons; Handbook of Experimental Immunology (ed. D.M. Weir and C.C. Blackwell); Gene Transfer Vectors for Mammalian Cells (ed. J.M. Miller and M.P. Calos, 1987); PCR: The Polymerase Chain Reaction, (ed. Mullis et al., 1994); Current Protocols in Immunology (ed. J.E. Coligan et al., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (ed. D. Catty., IRL Press, 1988-1989); Monoclonal Antibodies: A Practical Approach (ed. P. Shepherd and C. Dean, Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane, Cold Spring Harbor Laboratory Press, 1999); The Antibodies (ed. M. Zanetti and J. D. Capra, Harwood Academic Publishers, 1995); and Cancer: Principles and Practice of Oncology (V.T. DeVita et al., J.B. Lippincott Company, 1993).

I. Definitions

Unless otherwise indicated, the technical and scientific terms used in the present description have the meanings well known to the ordinary specialist in the field in which the present invention relates. In Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons, New York, N.Y. 1994 and March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 4th ed., John Wiley & Sons, New York, N.Y. 1992, a general interpretation of many of the concepts used in this application is provided for those skilled in the art. All references cited herein, including patent applications and patent publications, are hereby incorporated by reference in their entirety.

For the purposes of interpreting this specification, the following terms are to be used and, where possible, terms used in the singular also include their use in the plural and vice versa. It should be obvious that the terminology used in the present description is presented only for the purpose of describing specific embodiments of the invention and is not intended to limit its scope. In the event that any of the following definitions conflicts with those in any document incorporated by reference in the specification, the following definition shall be used.

For the purposes of this specification, "acceptor human framework" means a framework that contains the amino acid sequence of a light chain variable domain (VL) framework or a heavy chain variable domain (VH) framework derived from a human immunoglobulin framework or a human consensus framework, indicated below. An acceptor human framework region "derived from" a human immunoglobulin framework region or a human consensus framework region may have the same amino acid sequence or may contain alterations in the amino acid sequence. In some embodiments, the number of amino acid changes is 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less. In some embodiments, the sequence of the human VL acceptor framework is identical to the sequence of the human immunoglobulin VL framework or the sequence of the human consensus framework.

The concept of "affinity" refers to the total strength of all non-covalent interactions between one binding site of a molecule (eg, antibodies) and its binding partner (eg, antigen). As used herein, unless otherwise indicated, "binding affinity" refers to the intrinsic binding affinity of a molecule, reflecting a 1:1 interaction between the components of a binding pair (eg, antibody and antigen). The affinity of a molecule X for its partner Y can generally be characterized by the dissociation constant (Kd). Affinity can be assessed using conventional methods known in this field, including those presented in the present description. Specific options for measuring binding affinity are provided below by way of illustration.

An "affinity matured" antibody means an antibody with one or more changes in one or more hypervariable regions (HVRs) compared to a parent antibody that does not contain said changes, said changes leading to an increase in the affinity of the antibody for the antigen.

The terms "anti-myostatin antibody" and "antibody that binds to myostatin" refer to an antibody that has the ability to bind to myostatin with sufficient affinity to enable the antibody to be used as a diagnostic and/or therapeutic agent that targets myostatin. . In one embodiment, the level of binding of an anti-myostatin antibody to an unrelated non-myostatin protein is less than about 10% of the binding of the antibody to myostatin as measured by, for example, radioimmunoassay (RIA). In some embodiments, the antibody that binds to myostatin has a dissociation constant (Kd) of 1 μM or less, 100 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0.01 nM, or less than or 0.001 nM or less (for example, 10 ^-8 M or less, for example, from 10 ^-8 M to 10 ^-13 M, for example, from 10 ^-9 M to 10 ^-13 M). In some embodiments, the anti-myostatin antibody binds to a myostatin epitope that is conserved for myostatin from different species.

In the context of the present description, the term "antibody" is used in its broadest sense and refers to various structures of antibodies, including (but not limited to) monoclonal antibodies, polyclonal antibodies, multispecific antibodies (for example, bispecific antibodies) and antibody fragments, provided that they have the desired antigen-binding activity.

The term "antibody fragment" refers to a molecule, other than an intact antibody, that contains a portion of the intact antibody that binds to the antigen to which the intact antibody binds. Examples of antibody fragments include (but are not limited to) Fv, Fab, Fab', Fab'-SH, F(ab') ₂ ; dimeric antibodies (diabodi); linear antibodies; single chain antibody molecules (eg scFv); and multispecific antibodies formed from antibody fragments.

The term "antibody that binds to the same epitope" as a reference antibody refers to an antibody that blocks the binding of a reference antibody to its antigen when assessed by a competition assay (competitive assay) and/or, conversely, a reference antibody. the antibody blocks the binding of the antibody to its antigen when assessed by a competitive assay. An example of competitive analysis is presented in the present description.

The term "chimeric" antibody refers to an antibody in which part of the heavy and/or light chain is from a particular source or species, and the remainder of the heavy and/or light chain is from a different source or species.

The term "class" of an antibody refers to the type of constant domain or constant region that/which is included in its heavy chain. Five main classes of antibodies are known: IgA, IgD, IgE, IgG and IgM, and some of them can be further divided into subclasses (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant domains that correspond to the different immunoglobulin classes are referred to as alpha, delta, epsilon, gamma, and mu (α, δ, ε, γ, and μ), respectively.

The term "cytotoxic agent" in the context of the present description refers to a substance that inhibits or interferes with cellular function and/or causes cell death or destruction. Cytotoxic agents include (but not limited to) radioactive isotopes (for example, At ²¹¹ , I ¹³¹ , I ¹²⁵ , Y ⁹⁰ , Re ¹⁸⁶ , Re ¹⁸⁸ , Sm ¹⁵³ , Bi ²¹² , P ³² , Pb ²¹² and Lu radioactive isotopes); chemotherapeutic agents or drugs (eg, methotrexate, adriamycin, vinca alkaloids (vincristine, vinblastine, etoposide ), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents); growth inhibitory agents; enzymes and fragments thereof such as nucleolytic enzymes; antibiotics; toxins such as small molecular weight toxins or enzymatically active bacterial, fungal, plant or of animal origin, including fragments and/or variants thereof; and various antineoplastic or anticancer agents specified e below.

The term "effector functions" refers to those biological activities that are associated with the Fc region of the antibody, which vary depending on the isotype of the antibody. Examples of antibody effector functions include: C1q binding and complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-mediated cell-dependent cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (eg B cell receptor) and activation of B cells.

The term "effective amount" of an agent, for example, a pharmaceutical composition, means an amount effective in doses and for a period of time necessary to achieve the desired therapeutic or prophylactic result.

The term "epitope" includes any determinant that has the ability to bind an antibody. An epitope is a region of an antigen that binds to an antibody whose target is the antigen and includes specific amino acids that are in direct contact with the antibody. Epitope determinants may include reactive surface groups of molecules such as amino acids, sugar side chains, phosphoryl or sulfonyl groups, and may have specific three-dimensional structure characteristics and/or specific charge characteristics. In general, antibodies specific for a particular target antigen should preferentially recognize an epitope on the target antigen in a complex mixture of proteins and/or macromolecules.

"Fc receptor" or "FcR" means a receptor that binds to the Fc region of an antibody. In some embodiments, the FcR is a native human FcR. In some embodiments, the FcR is a receptor that binds to an IgG-type antibody (gamma receptor) and includes receptors of the subclass Fc-gamma RI, Fc-gamma RII, and Fc-gamma RIII, including allelic variants and resulting alternative splicing forms these receptors. Fc-gamma RII receptors include Fc-gamma RIIA ("activating receptor") and Fc-gamma RIIB ("inhibiting receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains. Activating receptor Fc-gamma RIIA contains immunoreceptor tyrosine-based activating motifs (ITAM) in its cytoplasmic domain. Inhibitory receptor Fc-gamma RIIB contains an immunoreceptor tyrosine receptor (ITIM) inhibitory motif in its cytoplasmic domain (see, for example, Daeron, Annu. Rev. Immunol. 15, 1997, pp. 203-234). An overview of FcR information is provided, for example, by Ravetch and Kinet, Annu. Rev. Immunol. 9, 1991, pp. 457-492; Capel et al., Immunomethods 4, 1994, pp. 25-34 and de Haas et al., J. Lab. Clin. Med. 126, 1995, pp. 330-341. Under the term "FcR", specified in the present description, fall and other FcR, including those that will be identified in the future.

The term "Fc receptor" or "FcR" also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgG to the embryo (Guyer et al., J. Immunol. 117, 1976, pp. 587-593 and Kim et al., J Immunol 24, 1994, 2429-2434) and regulates immunoglobulin homeostasis. Methods for measuring binding to FcRn are known (see, for example, Ghetie and Ward., Immunol. Today 18 (12), 1997, pp. 592-598; Ghetie et al., Nature Biotechnology 15 (7), 1997, pp. 637 -640 Hinton et al., J Biol Chem 279 (8) 2004 6213-6216 WO 2004/92219 (Hinton et al.) In vivo binding to human FcRn and human serum half-life An FcRn having a high binding affinity for polypeptides can be evaluated, for example, in transgenic mice or in transfected human cell lines expressing human FcRn or in primates administered polypeptides with a variant Fc region.WO 2000/42072 (Presta) describes antibody variants with increased or decreased ability to bind to FcR (see, for example, also Shields et al., J. Biol. Chem. 9(2), 2001, pp. 6591-6604).

The term "Fc region" in the context of the present description refers to the C-terminal region of the heavy chain of an immunoglobulin, which contains at least part of the constant region. The term includes native sequence Fc regions and variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) or glycine-lysine (residues 446-447) of the Fc region may or may not be present. Unless specifically stated otherwise, the numbering of amino acid residues in the Fc region or constant region follows the EU numbering system, which is also referred to as the EU index, as described by Kabat E.A. et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

The term "containing an Fc region antibody" refers to an antibody that contains an Fc region. The C-terminal lysine (residue 447 according to the EU numbering system) or the C-terminal glycine-lysine (residues 446-447) of the Fc region can be removed, for example, during purification of the antibody or by recombinant construction of the nucleic acid encoding the antibody. Thus, a composition that contains an antibody having an Fc region of the present invention may contain an antibody with G446-K447, with G446 and without K447, with all G446-K447 deleted, or a mixture of the three types of antibodies described above.

"Framework region" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The variable domain FR typically consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences typically have the following location in the VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

The terms "full-length antibody", "intact antibody", and "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to the native antibody structure, or having heavy chains that contain the Fc region described herein.

A "functional Fc region" has an "effector function" inherent in a native sequence Fc region. Examples of "effector functions" include C1q binding; (CDC); Fc receptor binding; ADCC; phagocytosis; downregulation of cell surface receptors (eg B cell receptor; BCR), etc. These effector functions generally require the Fc region to be combined with a binding domain (eg, an antibody variable domain) and can be assessed using various assays, such as those provided in the "Definitions" section of this specification.

The terms "host cell", "host cell line" and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including progeny of said cells. Host cells include "transformants" and "transformed cells", which include the primary transformed cell and its progeny, regardless of the number of passages. The offspring may not be completely identical in nucleic acid composition to the parent cell, but may contain mutations. Under the scope of the invention falls mutant progeny, which has the same function or biological activity, which is found as a result of screening or selected from the original transformed cell.

A "human antibody" is an antibody having an amino acid sequence that corresponds to that of an antibody produced in the human body or human cells, or derived from a non-human source that uses a human antibody spectrum or other human antibody-encoding sequences. Specifically excluded from this definition of a human antibody is a humanized antibody containing non-human antigen-binding residues.

A "human consensus framework region" is a framework region that includes the most frequently occurring amino acid residues in selected human immunoglobulin VL or VH framework sequences. Typically, the choice of human immunoglobulin VL or VH sequences is made from a subset of variable domain sequences. Generally, a subgroup of sequences is that described by Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed., NIH Publication 91-3242, Bethesda MD, vols. 1-3, 1991. In one of the VL embodiments, the subgroup is the kappa I subgroup according to Kabat et al., supra. In one embodiment of the invention regarding VH, the subgroup is subgroup III according to Kabat et al., supra.

The term "humanized" antibody refers to a chimeric antibody that contains amino acid residues from non-human HVRs and amino acid residues from human FRs. In some embodiments, a humanized antibody may comprise substantially all of at least one, and typically two, variable domains in which all or substantially all of the HVRs (e.g., CDRs) correspond to regions of the non-human antibody and all or substantially all of the FRs correspond to regions human antibody. A humanized antibody may optionally contain at least a portion of an antibody constant region derived from a human antibody. The term "humanized form" of an antibody, such as a non-human antibody, refers to an antibody that has been humanized.

The term "hypervariable region" or "HVR" in the context of the present description refers to each of the regions of the variable domain of an antibody, the sequences of which are hypervariable ("complementarity determining regions" or "CDRs") and/or form loops of a certain structure ("hypervariable loops") , and/or contain antigen-contacting residues ("antigen-contacts"). Typically, antibodies contain six HVRs; three in VH (H1, H2, H3) and three in VL (LI, L2, L3). As used herein, examples of HVRs include (a) hypervariable loops comprising amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196, 1987, pp. 901-917); (b) CDRs including amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2) and 95-102 (H3) ( Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, 1991, NIH publication 91-3242); (c) antigen contact regions comprising amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2) and 93-101 ( H3) (MacCallum et al., J. Mol. Biol. 262, 1996, pp. 732-745); and (d) combinations of residues specified in subparagraphs (a), (b) and/or (c), including amino acid residues HVR 46-56 (L2), 47-56 (L2), 48-56 (L2), 49 -56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3) and 94-102 (H3).

Unless specifically stated otherwise, residues in HVR and other residues in the variable domain (eg, residues in FR) are numbered according to Kabat et al., supra.

An "immunoconjugate" is an antibody conjugated to one or more heterologous molecule(s), including (but not limited to) a cytotoxic agent.

"Individual" or "subject" is a mammal. Mammals include, but are not limited to, domesticated animals (for example, cows, sheep, cats, dogs, and horses), primates (for example, humans and non-human primates, such as monkeys), rabbits, and rodents (for example, mice and rats). In some embodiments of the invention, the individual or subject is a human.

An "isolated" antibody is one that has been separated from a component of its natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity as determined by, for example, electrophoretic assays (such as, for example, SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis) or chromatographic assays (such as, for example, as ion exchange chromatography or reverse phase HPLC). For a review of methods for assessing antibody purity, see, for example, Flatman et al., J. Chrom. In 848, 2007, pp. 79-87.

An "isolated" nucleic acid is a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in a cell that normally includes a nucleic acid molecule, but in which the nucleic acid molecule is present outside the chromosome or in which its localization on the chromosome differs from its naturally occurring localization on the chromosome.

The term “isolated nucleic acid encoding an anti-myostatin antibody” refers to one or more nucleic acid molecules encoding heavy and light chains of an antibody (or fragments thereof), including the specified molecule(s) in one vector or in different vectors, and said nucleic acid molecule(s) present(s) at one or more positions in the host cell.

The term "monoclonal antibody" in the context of the present description refers to an antibody obtained from a population of substantially homogeneous antibodies, i. individual antibodies in a population are identical and/or bind to the same epitope, with the exception of possible variants of antibodies, for example, containing mutations that occur naturally or arise during the production of a preparation of a monoclonal antibody, these variants are usually present in minor quantities. Unlike polyclonal antibody preparations, which typically include different antibodies to different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed against a single antigen determinant. Thus, the adjective "monoclonal" defines the characteristics of an antibody, characterizing it as derived from a substantially homogeneous population of antibodies, rather than being engineered to meet the requirements for producing an antibody by any particular method. For example, monoclonal antibodies that can be used according to the present invention can be created using various technologies, including (but not limited to) the hybridoma method, recombinant DNA methods, phage display methods, and methods based on the use of transgenic animals, which contain all or a portion of human immunoglobulin loci, these methods and other exemplary methods for producing monoclonal antibodies are provided herein.

The term "naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (eg, a cytotoxic moiety) or a radioactive label. The naked antibody may be present in the pharmaceutical composition.

The term "native antibodies" refers to naturally occurring immunoglobulin molecules with different structures. For example, native IgG antibodies are heterotetrameric glycoproteins with a molecular weight of approximately 150,000 Da, consisting of two identical light chains and two identical heavy chains linked by disulfide bridges. From the N-terminus to the C-terminus, each heavy chain has a variable region (VH), which is also referred to as a heavy chain variable domain or a heavy chain variable domain, followed by three constant domains (CH1, CH2 and CH3). Similarly, from the N-terminus to the C-terminus, each light chain has a variable region (VL), also referred to as a variable light domain or a light chain variable domain, followed by a light chain constant domain (CL). The light chain of an antibody, depending on the amino acid sequence of its constant domain, can belong to one of two types, which are kappa (κ) and lambda (λ).

A "native sequence Fc region" contains an amino acid sequence identical to that of a naturally occurring Fc region. The native sequence of human Fc regions includes the native sequence of the human IgG1 Fc region (non-A and A allotypes); native sequence of the Fc region of human IgG2; the native sequence of the human IgG3 Fc region and the native sequence of the human IgG4 Fc region, as well as their naturally occurring variants.

The term "package leaflet" refers to the instructions that are usually included in the commercial packaging of therapeutic products, containing information about the indications, use, dose, route of administration, combination therapy, contraindications and / or precautions associated with the use of these therapeutic products.

"Percent (%) Amino Acid Sequence Identity" relative to a reference polypeptide is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in the reference polypeptide, after sequence alignment and, if necessary, the introduction of gaps to achieve the maximum percent sequence identity, and without considering any conservative substitutions as a component in assessing sequence identity. Comparative analysis for purposes of determining percent amino acid sequence identity can be performed in various ways known in the art using, for example, publicly available computer programs such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) or GENETYX® (Genetyx Co. ., Ltd.) Those skilled in the art can determine appropriate parameters for sequence alignment, including any algorithms necessary to achieve maximum alignment over the entire length of the compared sequences. The sequence comparison computer program ALIGN-2 is copyrighted by Genentech, Inc. and the source code has been provided in the U.S. user documentation. Copyright Office, Washington D.C., 20559, where it is registered as U.S. Copyright Registration No. TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, CA. California, or it can be compiled from source code. The ALIGN-2 program can be compiled for use on the UNIX operating system, including Digital UNIX V4.0D. All parameters required for sequence comparison are set by ALIGN-2 and should not be changed.

In situations in which ALIGN-2 is used to compare amino acid sequences, the % amino acid sequence identity of a given amino acid sequence A compared to, with, or relative to a given amino acid sequence B (which can alternatively be referred to as a given amino acid sequence A that has or contains a specific % Amino Acid Sequence Identity compared to, with, or relative to a given Amino Acid Sequence B) is calculated as follows: 100 x Fraction X/Y where X is the number of amino acid residues determined to be identical matches when assessed using the ALIGN-2 sequence comparison software , where the program compared A and B, and where Y denotes the total number of amino acid residues in B. It should be obvious that if the length of the amino acid sequence A is not equal to the length of the amino acid sequence B, then % The % Amino Acid Sequence Identity of A to B cannot be equal to % Amino Acid Sequence Identity of B to A. Unless specifically stated otherwise, all % Amino Acid Sequence Identities referred to herein were obtained using the ALIGN-2 computer program described in the previous paragraph.

The term "pharmaceutical composition" refers to a preparation that is in such a form that it provides the biological activity of the active substance included in its composition, which should be effective, and which does not contain additional components that have unacceptable toxicity for the individual to whom the composition should be administered.

A "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical composition, other than the active ingredient, that is non-toxic to an individual. Pharmaceutically acceptable carriers include, but are not limited to, a buffer, excipient, stabilizer, or preservative.

The term "myostatin" as used herein can refer to any native myostatin from any vertebrate animal, including mammals such as primates (eg humans) and rodents (eg mice and rats). Unless specifically stated otherwise, the term "myostatin" refers to a human myostatin protein having the amino acid sequence shown in SEQ ID NO: 1 and containing the terminal human myostatin propeptide domain shown in SEQ ID NO: 75 or 78. The term refers to " full-length" unprocessed myostatin, as well as to the form of myostatin, which is formed as a result of processing in the cell. The term also includes naturally occurring variants of human myostatin, such as splicing variants or allelic variants. The amino acid sequence of human myostatin (promyostatin) is shown in SEQ ID NO: 1. The amino acid sequence of exemplary human myostatin growth factor C-terminal domain is shown in SEQ ID NO: 2. The amino acid sequence of exemplary human myostatin N-terminal propeptide domain shown in SEQ ID NO: 75 or 78. Active mature myostatin is a disulfide-linked homodimer consisting of two C-terminal growth factor domains. Inactive latent myostatin is a non-covalently associated complex of two propeptides and mature myostatin. As described herein, the antibodies of the invention bind to inactive latent myostatin but do not bind to mature active myostatin homodimer. In some embodiments, the antibodies of the invention bind to an epitope within amino acids 21-100 of the myostatin propeptide (SEQ ID NO: 78) but do not bind to the mature active myostatin homodimer. The amino acid sequence of exemplary cynomolgus monkey myostatin and mouse myostatin (promyostatin) are shown in SEQ ID NOS: 3 and 5, respectively. The amino acid sequence of the exemplary C-terminal domain of monkey cynomolgus myostatin growth factor and mouse myostatin is shown in SEQ ID NOS: 4 and 6, respectively. The amino acid sequence of the exemplary N-terminal propeptide domain of monkey cynomolgus myostatin and mouse myostatin is shown in SEQ ID NOs: 76 or 79 and 77 or 80, respectively. GDF-11 (BMP-11) is a closely related molecule to myostatin, both of which are members of the TGF-beta superfamily. Similar to myostatin, GDF11 is synthesized first as a precursor polypeptide and then cleaved into an N-terminal prodomain and a C-terminal mature GDF11. The amino acid sequence of human GDF11 (precursor) is shown in SEQ ID NO: 81. The amino acid sequence of C-terminal mature human GDF11 is shown in SEQ ID NO: 82. The amino acid sequence of the N-terminal prodomain of human GDF11 is shown in SEQ ID NO: 83 or 84. sequence SEQ ID NO: 1, 3, 5, 78, 79, 80, 81 and 84 include a signal sequence. It corresponds to amino acids 1-24, and they are removed during processing in the cell.

In the context of the present description, the term "treatment" (and its grammatical variations, such as "treat" or "process of treatment") refers to a clinical intervention with the aim of changing the natural course of the disease in the individual being treated, and it can be carried out either for prevention or during the development of clinical pathology. The desired actions of treatment are (but not limited to) prevention of the onset or recurrence of the disease, relief of symptoms, reduction of any direct or indirect pathological consequences of the disease, prevention of metastases, reduction in the rate of disease progression, alleviation or temporary amelioration of the disease state, and remission or improvement in prognosis. In some embodiments, the antibodies of the invention are used to delay the development of a disease or slow the progression of a disease.

The term "variable region" or "variable domain" refers to the heavy or light chain domain of an antibody that is involved in the binding of an antibody to an antigen. The heavy chain and light chain variable domains (VH and VL, respectively) of a native antibody generally have similar structures, with each domain containing four conserved framework regions (FR) and three hypervariable regions (HVR) (see, e.g., Kindt and et al., Kuby Immunology, 6th ed., W. H. Freeman and Co., 2007, p. 91). A single VH or VL domain may be sufficient to provide antigen binding specificity. In addition, antibodies that bind to a particular antigen can be isolated using the VH or VL domain from an antibody that binds to the antigen to screen for a library of complementary VL or VH domains, respectively (see, e.g., Portolano et al. , J. Immunol 150, 1993, pp. 880-887; Clarkson et al., Nature 352, 1991, pp. 624-628).

"Fc-region variant" contains an amino acid sequence that differs from the native Fc-region sequence due to the presence of at least one amino acid modification (change), preferably one or more amino acid(s) substitution(s). Preferably, the Fc region variant has at least one amino acid substitution relative to the native Fc region sequence of the parent polypeptide, e.g., from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in the native Fc region sequence. or in the Fc region of the parent polypeptide. In the context of the present description, the Fc region variant should preferably have at least about 80% homology with the native sequence of the Fc region and/or Fc region of the parent polypeptide, and most preferably at least about 90% homology with it, more preferably at least about 95% homology with it.

The term "vector" in the context of the present description refers to a nucleic acid molecule that has the ability to reproduce another nucleic acid with which it is associated. The concept includes a vector in the form of a self-replicating nucleic acid structure, as well as a vector included in the genome of the host cell into which it is introduced. Some vectors have the ability to control the expression of the nucleic acids to which they are operably linked. These vectors in the context of the present description are referred to as "expression vectors".

II. Compositions and Methods

One of the objects of the invention are, in particular, antibodies to myostatin and their uses. Some embodiments of the invention are antibodies that bind to myostatin. The antibodies of the invention can be used, for example, to diagnose or treat muscle wasting syndrome.

One of the objects of the invention are, in particular C5, polypeptides containing variants of the Fc regions, and their uses. One of the embodiments of the invention are polypeptides containing variants of the Fc regions with increased Fc-gamma RIIb-binding activity. Another embodiment of the invention are polypeptides containing variant Fc regions with increased pI. In specific embodiments of the invention, the polypeptides of the invention are antibodies. Polypeptides containing a variant Fc region of the invention can be used, for example, to diagnose or treat a disease.

A. Examples of Anti-Myostatin Antibodies and Polypeptides Containing Variant Fc Regions

One of the objects of the invention are isolated antibodies that bind to myostatin. In some embodiments, an anti-myostatin antibody of the present invention binds to latent myostatin. In other embodiments, an anti-myostatin antibody of the present invention binds to a myostatin propeptide (human: SEQ ID NO: 75 or 78; cynomolgus monkey: SEQ ID NO: 76 or 79; mouse: SEQ ID NO: 77 or 80) . In other embodiments, the antibody binds to an epitope in a fragment consisting of amino acids 21-100 of the myostatin propeptide (SEQ ID NO: 78). The propeptide is included in latent myostatin as one of the components, as described above. In some embodiments, an anti-myostatin antibody of the present invention inhibits myostatin activation. In some embodiments, the anti-myostatin antibody blocks the release of mature myostatin from latent myostatin. Mature myostatin is known to be released through proteolytic and non-proteolytic processes from latent myostatin. An anti-myostatin antibody of the present invention can block the proteolytic and/or non-proteolytic release of mature myostatin from latent myostatin. In some embodiments, the anti-myostatin antibody blocks the proteolytic cleavage of latent myostatin. In some embodiments, the anti-myostatin antibody blocks the access of a protease to latent myostatin (primarily the proteolytic cleavage site (Arg98-Asp99) of latent myostatin). In other embodiments, the protease may be a member of the BMP1/TLD metalloprotease family, such as BMP1, TED, tolloid-like protein-1 (TLL-1), or tolloid-like protein-2 (TLL-2). In another embodiment, the anti-myostatin antibody blocks the non-proteolytic release of mature myostatin from latent myostatin. In the context of the present description, non-proteolytic release means the spontaneous release of mature myostatin from latent myostatin, which is not accompanied by proteolytic cleavage of latent myostatin. Ileproteolytic release includes, for example, the release of mature myostatin by incubation of latent myostatin, for example at 37° C., in the absence of a protease that cleaves latent myostatin. In some embodiments, an anti-myostatin antibody of the present invention does not bind to mature myostatin. In some embodiments, the anti-myostatin antibody binds to the same epitope as the antibody shown in Table 2a. In some embodiments, the anti-myostatin antibody competes for binding to latent myostatin with the antibody shown in Table 2a. In additional embodiments, the anti-myostatin antibody competes for binding to latent myostatin, with an antibody that contains the VH and VL pair shown in Table 2a. In some embodiments, the anti-myostatin antibody competes for binding to the amino acid 21-100 fragment of the myostatin propeptide (SEQ ID NO: 78) with the antibody shown in Table 2a. In other embodiments, the anti-myostatin antibody binds to the same epitope as the antibody shown in Table 11a or 13. In some embodiments, the anti-myostatin antibody competes for binding to latent myostatin with the antibody shown in Table 11a or 13. B In some embodiments, the anti-myostatin antibody competes for binding to the amino acid 21-100 fragment of the myostatin propeptide (SEQ ID NO: 78) with the antibody shown in Table 11a or 13.

In some embodiments, an anti-myostatin antibody of the present invention binds to latent myostatin and inhibits myostatin activation. In other embodiments, the antibody: (a) blocks the release of mature myostatin from latent myostatin; (b) blocks the proteolytic release of mature myostatin; (c) blocks the spontaneous release of mature myostatin or (d) does not bind to mature myostatin; or binds to an epitope within a fragment consisting of amino acids 21-100 of the myostatin propeptide (SEQ ID NO: 78). In other embodiments, the antibody competes for binding to latent myostatin or binds to the same epitope as an antibody that contains the VH and VL pair shown in Tables 2a, 11a, or 13. In other embodiments, the antibody binds to latent myostatin with higher affinity at neutral pH (eg pH 7.4) than at acidic pH (eg pH 5.8). In other embodiments, the antibody is (a) a monoclonal antibody, (b) a human, humanized, or chimeric antibody; (c) a full-length IgG-type antibody; or (d) an antibody fragment that binds to latent myostatin or myostatin propeptide.

In another embodiment, the anti-myostatin antibody of the present invention does not bind to GDF11 (growth factor 11). In some embodiments, an anti-myostatin antibody of the present invention does not inhibit GDF11 activation. In some embodiments, the anti-myostatin antibody does not block the release of mature GDF11 from latent GDF11. The anti-myostatin antibody of the present invention does not block either proteolytic or non-proteolytic release of mature GDF11 from latent GDF11. In some embodiments, the anti-myostatin antibody does not block proteolytic cleavage of latent GDF11. In some embodiments, the anti-myostatin antibody does not block protease access to latent GDF11 (primarily the proteolytic cleavage site of latent GDF11). In other embodiments, the protease may be a metalloprotease from the BMP1/TLD family, such as BMP1, TED, tolloid-like protein-1 (TLL-1), or tolloid-like protein-2 (TLL-2). Under non-proteolytic release in the context of the present description refers to the spontaneous release of Mature GDF11 from latent GDF11, which is not carried out through proteolytic cleavage of latent GDF11. Ileproteolytic release includes, for example, the release of mature GDF11 by incubation of latent GDF11, for example, at 37°C in the absence of a protease that cleaves latent GDF11. Most of the anti-myostatin antibodies known to date are not specific for myostatin. These antibodies have high affinity for other members of the TGF-beta superfamily, such as GDF11, and neutralize their biological activity. GDF11 plays an important role in the process of embryogenesis and is responsible for the homeotic transformation of the axial skeleton. Mice homozygous for the "off" GDF11 died in the perinatal period, mice with one copy of the wild type GDF11 gene were viable, but had skeletal defects. Since GDF11 plays an important role in the process of embryogenesis, an antagonist that inhibits GDF11 theoretically carries safety risks, which can manifest itself as either toxicity to treated patients or reproductive toxicity, for example, to women of childbearing potential. Thus, there is a need for specific inhibition of myostatin activity in the treatment of myostatin-associated disorders in which an increase in mass, size, strength, etc. is desired. muscles, especially in women of childbearing potential.

Another object of the invention are antibodies to myostatin with pH-dependent binding characteristics. In the context of the present description, the term "pH-dependent binding" means that the antibody has a "reduced ability to bind to myostatin at acidic pH compared to its binding at neutral pH" (both terms can be used interchangeably for the purposes of this description). For example, antibodies with "pH dependent binding characteristics" include antibodies that bind to myostatin with higher affinity at neutral pH than at acidic pH. In some embodiments, the antibodies of the present invention bind to myostatin with an affinity of at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10,000 or more times higher at neutral pH than at acidic pH. In some embodiments, antibodies bind to myostatin (eg, latent myostatin or myostatin propeptide) with higher affinity at pH 7.4 than at pH 5.8. In other embodiments, the antibodies bind to myostatin with an affinity of at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10,000 or more times higher at pH 7.4 than at pH 5.8.

When the antigen is a soluble protein, binding of the antibody to the antigen may result in an extended plasma half-life of the antigen (i.e., reduced clearance of the antigen from plasma), since the antibody may have a longer plasma half-life than the antigen itself, and can serve as a carrier for an antigen. This is a consequence of the recycling of the antigen-antibody complex by FcRn through the endosomal pathway in the cell (Roopenian, Nat. Rev. Immunol. 7(9), 2007, pp. 715-725). However, an antibody with pH-dependent binding characteristics that binds to its antigen in a neutral extracellular environment, while releasing the antigen into acidic endosomal compartments upon entry into cells, would be expected to have advantageous neutralization and clearance properties of the antigen relative to the copy that binds. in a pH independent manner (Igawa et al., Nature Biotechnol. 28(11), 2010, pp. 1203-1207; Devanaboyina et al., mAbs 5(6), 2013, pp. 851-859; WO 2009/125825) .

For the purposes of this specification, the "affinity" of an antibody to myostatin is expressed in terms of the KD of the antibody. The KD of an antibody means the dissociation equilibrium constant of the antibody-antigen interaction. The higher the KD value, which characterizes the binding of an antibody to its antigen, the weaker its binding affinity for the specified specific antigen. Thus, in the context of the present description, the concept of "higher affinity at neutral pH than at acidic pH" (or the equivalent concept of "pH-dependent binding") means that the KD, which characterizes the binding of an antibody to myostatin at acidic pH, is higher than the KD , which characterizes the binding of antibodies to myostatin at neutral pH. For example, in the context of the present invention, an antibody is considered to bind to myostatin with higher affinity at neutral pH than at acidic pH if the KD characterizing the binding of the antibody to myostatin at acidic pH is at least 2 times higher than the KD characterizing binding of the antibody to myostatin at neutral pH. Thus, the present invention includes antibodies whose binding to myostatin at acidic pH is characterized by a KD value that is at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10,000 or more times the KD of myostatin binding at neutral pH. In another embodiment, the KD value at neutral pH may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less. In another embodiment of the invention, the value of KD at acidic pH may be 10 ^-9 M, 10 ^-8 M, 10 ^-7 M, 10 ^-6 M or more.

In other embodiments, an antibody is said to bind to myostatin (e.g., latent myostatin or myostatin propeptide) with higher affinity at neutral pH than at acidic pH if the KD value of antibody binding to myostatin at pH 5.8 is at least 2 times higher than the KD, which characterizes the binding of the antibody to myostatin at pH 7.4. In some embodiments of the invention, the binding of the proposed antibodies to myostatin at pH 5.8 is characterized by a KD value that is at least 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times the KD characterizing the binding of the antibody to myostatin at pH 7.4. In another embodiment, the KD value of the antibody at pH 7.4 may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less. In another embodiment, the KD value of the antibody at pH 5.8 may be 10 ^-9 , 10 ^-8 , 10 ^-7 , 10 ^-6 M or more.

The binding characteristics of an antibody to a particular antigen can also be expressed as the kd of an antibody. The kd value of an antibody denotes the dissociation rate constant of an antibody relative to a particular antigen and is expressed as reciprocals of a second (ie, s ⁻¹ ). An increased kd value means weaker binding of the antibody to its antigen. Thus, the present invention includes antibodies whose binding to myostatin has a higher kd value at acidic pH than at neutral pH. The present invention includes antibodies whose binding to myostatin at acidic pH is characterized by a kd value that is at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 , 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times the kd characterizing the binding of the antibody to myostatin at neutral pH. In another embodiment, the kd value of the antibody at neutral pH may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at acidic pH may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more. The invention also includes antibodies whose binding to myostatin (eg, latent myostatin or myostatin propeptide) has a higher kd value at pH 5.8 than at pH 7.4. The invention includes antibodies whose binding to myostatin at pH 5.8 is characterized by a kd value that is at least 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times the kd characterizing the binding of the antibody to myostatin at pH 7.4. In another embodiment, the kd value of the antibody at pH 7.4 may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at pH 5.8 may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more.

In some cases, "reduced ability to bind to myostatin at acidic pH compared to the ability to bind at neutral pH" is expressed as the ratio of the KD value, which characterizes the binding of the antibody to myostatin at acidic pH, and the KD value, which characterizes the binding of the antibody to myostatin at neutral pH ( or vice versa). For example, an antibody may be considered to have "reduced ability to bind to myostatin at acidic pH compared to the ability to bind at neutral pH" for the purposes of the present invention if the antibody's acid/neutral pH KD ratio is 2 or more. In some embodiments, the KD ratio at pH 5.8/pH 7.4 for an anti-myostatin antibody of the present invention is 2 or more. In some embodiments, the acid/neutral pH KD ratio for an antibody of the present invention may be 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the KD value of the antibody at neutral pH may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 - ¹¹ , 10 ^-12 M or less. In another embodiment, the KD value of the antibody at acidic pH may be 10 ^-9 , 10 ^-8 , 10 ^-7 , 10 ^-6 M or more. In other instances, an antibody may be considered to have "reduced ability to bind to myostatin (e.g., latent myostatin) at acidic pH compared to binding at neutral pH" if the antibody has a KD ratio at pH 5.8/pH 7.4, component 2 or more. In some exemplary embodiments of the invention, the KD ratio at pH 5.8/pH 7.4 for an antibody may be 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 , 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the KD value of the antibody at pH 7.4 may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less. In another embodiment, the KD value of the antibody at pH 5.8 may be 10 ^-9 , 10 ^-8 , 10 ^-7 , 10 ^-6 M or more.

In some cases, "reduced ability to bind to myostatin at acidic pH compared to the ability to bind at neutral pH" is expressed as the ratio of the kd value, which characterizes the binding of the antibody to myostatin at acidic pH, and the kd value, which characterizes the binding of the antibody to myostatin at neutral pH ( or vice versa). For example, an antibody may be considered to have "reduced ability to bind to myostatin at acidic pH compared to the ability to bind at neutral pH" for the purposes of the present invention if the antibody's acid/neutral pH kd ratio is 2 or more. In some embodiments, the kd ratio at pH 5.8/pH 7.4 for an anti-myostatin antibody of the present invention is 2 or more. In some embodiments, the acid/neutral pH kd ratio for an antibody of the present invention may be 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the kd value of the antibody at neutral pH may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at acidic pH may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more. In some exemplary embodiments of the invention, the ratio kd at pH 5.8/pH 7.4 for an antibody may be 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 , 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the kd value of the antibody at pH 7.4 may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at pH 5.8 may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more.

In the context of the present description, the term "acidic pH" means a pH from 4.0 to 6.5. The term "acidic pH" includes one of the following pH values 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 , 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6 .2, 6.3, 6.4 and 6.5. In specific aspects of the invention, "acidic pH" corresponds to 5.8.

In the context of the present description, the term "neutral pH" means a pH from 6.7 to about 10.0. The term "neutral pH" includes one of the following pH values 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6 , 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8 .9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, and 10.0. In specific aspects of the invention, "neutral pH" corresponds to 7.4.

The KD and kd values described herein can be determined using a biosensor based surface plasmon resonance method to characterize antibody-antigen interactions (see, for example, Example 7 herein). The KD values and kd values can be determined at 25°C or 37°C.

In some embodiments, an anti-myostatin antibody of the present invention binds to more than one species of myostatin. In other embodiments, the anti-myostatin antibody binds to human and non-human myostatin. In other embodiments, the anti-myostatin antibody binds to human, murine, and monkey myostatin (eg, cynomolgus monkeys, rhesus monkeys, marmosets, chimpanzees, or baboons).

In some embodiments, an anti-myostatin antibody of the present invention binds to latent myostatin from more than one species. In other embodiments, the anti-myostatin antibody binds to latent human and non-human myostatin. In other embodiments, the anti-myostatin antibody binds to latent human, mouse, and monkey myostatin.

In some embodiments, an anti-myostatin antibody of the present invention binds to a myostatin propeptide from more than one species. In other embodiments, the anti-myostatin antibody binds to a human and non-human myostatin propeptide. In other embodiments, the anti-myostatin antibody binds to human, mouse, and monkey myostatin propeptide.

Another aspect of the invention is an anti-myostatin antibody that forms an immune complex (ie, an antigen-antibody complex) with myostatin. In some embodiments, two or more anti-myostatin antibodies bind to two or more myostatin molecules to form an immune complex. This is possible because myostatin exists as a homodimer that contains two myostatin molecules and the antibody has two antigen binding sites. An anti-myostatin antibody may bind to the same epitope on the myostatin molecule or may bind to different epitopes on the myostatin molecule, most likely such an antibody is a bispecific antibody. In general, when two or more antibodies form an immune complex with two or more antigens, the resulting immune complex can bind strongly to Fc receptors present on cell surfaces due to avidity effects through the Fc regions of the antibodies in the complex and can then absorbed by cells with high efficiency. Thus, the above anti-myostatin antibody, which has the ability to form an immune complex containing two or more anti-myostatin antibodies and two or more molecules of myostatin, can lead to rapid clearance of myostatin from plasma in vivo due to strong binding to Fc receptors. due to the effects of avidity.

Furthermore, it is contemplated that an antibody with pH-dependent binding characteristics may have advantageous antigen-clearance-neutralizing properties over its copy that binds in a pH-independent manner (Igawa et al., Nature Biotech. 28(11), 2010, cc 1203-1207; Devanaboyina et al., mAbs 5 (6), 2013, pp. 851-859; WO 2009/125825). Thus, an antibody having the above two properties, i. an antibody having pH-dependent binding characteristics and forming an immune complex, which contains two or more antibodies with two or more antigens, should have even more advantageous properties regarding the accelerated elimination of antigens from plasma (WO 2013/081143).

Another object of the invention is an antibody to myostatin, which contains at least one, two, three, four, five or six HVRs selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55-57, 114 -115, 126; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60, 116-120, 127; (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64, 121, 128; (d) HVR-L1 which contains one of the amino acid sequences of SEQ ID NOs: 65-69, 122-124, 129; (e) HVR-L2 which contains one of the amino acid sequences of SEQ ID NOs: 70-72, 125, 130; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74, 131.

Another object of the invention is an antibody to myostatin, which contains at least one, two, three, four, five or six HVR selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55-57; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60; (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64; (d) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 65-69; (e) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74.

One of the objects of the invention is an antibody to myostatin, which contains at least one, two, three, four, five or six hypervariable regions (HVR) selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 114-115; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 116-120; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 121; (d) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 122-124; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 125; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74.

Another object of the invention is an antibody to myostatin, which contains at least one, two, three, four, five or six HVR selected from (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 114; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63; (d) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 122; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (e) an HVR-L3 which contains the amino acid sequence of SEQ ID NO: 74. Another aspect of the invention is an anti-myostatin antibody which contains at least one, two, three, four, five, or six HVRs selected from (a) HVRs -H1, which contains the amino acid sequence of SEQ ID NO: 114; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63; (d) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 123; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (e) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 74.

Another object of the invention is an antibody to myostatin, which contains at least one, two, three, four, five or six HVR selected from (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 126; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 127; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 128; (d) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 129; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 130; and (e) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 131.

One of the objects of the invention is an antibody that contains at least one, at least two or all three VH HVR sequences selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55-57, 114 -115, 126; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60, 116-120, 127; (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64, 121, 128. In one embodiment, the antibody comprises an HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64 , 121, 128. In another embodiment, the antibody comprises HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64, 121, 128, and HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73 -74, 131. In the following embodiment, the antibody comprises HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64, 121, 128, HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73- 74, 131, and HVR-H2, which contains one of the amino acid sequences of SEQ ID NOs: 58-60, 116-120, 127. In the following embodiment, the antibody contains (a) HVR-H1, which contains one of the amino acid sequences her SEQ ID NOs: 55-57, 114-115, 126; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60, 116-120, 127; and (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64, 121, 128.

One of the objects of the invention is an antibody that contains at least one, at least two or all three VH HVR sequences selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55-57; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60; and (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64. In one of the embodiments of the invention, the antibody contains HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64. In another embodiment, the antibody contains HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64, and HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73-74. In a further embodiment, the antibody comprises HVR-H3, which contains one of the amino acid sequences of SEQ ID NOs: 61-64, HVR-L3, which contains one of the amino acid sequences of SEQ ID NOs: 73-74, and HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60. In the following embodiment of the invention, the antibody contains (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55-57; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60; and (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64.

Another object of the invention is an antibody that contains at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 114-115; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 116-120; and (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 121. In one embodiment, the antibody contains HVR-H3, which contains the amino acid sequence of SEQ ID NO: 121. In another embodiment, the antibody contains HVR-H3 , which contains the amino acid sequence of SEQ ID NO: 121, and HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73-74. In a further embodiment, the antibody comprises HVR-H3, which contains the amino acid sequence of SEQ ID NO: 121, HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73-74, and HVR-H2, which contains one of the amino acid sequences SEQ ID NO: 116-120. In the following embodiment of the invention, the antibody contains (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 114-115; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 116-120; and (c) HVR-H3 which contains the amino acid sequence of SEQ ID NO: 121.

Another object of the invention is an antibody that contains at least one, at least two, or all three VH HVR sequences selected from (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 114; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58; and (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63. In one embodiment, the antibody contains HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63. In another embodiment, the antibody contains HVR-H3 , which contains the amino acid sequence of SEQ ID NO: 63, and HVR-L3, which contains the amino acid sequence of SEQ ID NO: 74. In the following embodiment, the antibody contains HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63, HVR-L3 , which contains the amino acid sequence of SEQ ID NO: 74, and HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58. In another embodiment, the antibody contains (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 114; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58; and (c) HVR-H3 which contains the amino acid sequence of SEQ ID NO: 63.

Another object of the invention is an antibody containing at least one, at least two or all three HVR VH sequences selected from (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 126; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 127; and (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 128. In one embodiment, the antibody comprises HVR-H3, which contains the amino acid sequence of SEQ ID NO: 128. In another embodiment, the antibody comprises HVR-H3 , which contains the amino acid sequence of SEQ ID NO: 128, and HVR-L3, which contains the amino acid sequence of SEQ ID NO: 131. In the following embodiment, the antibody contains HVR-H3, which contains the amino acid sequence of SEQ ID NO: 128, HVR-L3 , which contains the amino acid sequence of SEQ ID NO: 131, and HVR-H2, which contains the amino acid sequence of SEQ ID NO: 127. In another embodiment, the antibody contains (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 126; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 127; and (c) HVR-H3 which contains the amino acid sequence of SEQ ID NO: 128.

Another object of the invention is an antibody containing at least one, at least two or all three HVR VL sequences selected from (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 65-69,122-124, 129; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72, 125, 130; and (c) HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73-74, 131. In one embodiment, the antibody contains (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 65 -69.122-124, 129; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72, 125, 130; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74, 131.

Another object of the invention is an antibody containing at least one, at least two or all three HVR VL sequences selected from (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 65-69; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74. In one of the embodiments of the invention, the antibody contains (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 65-69; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74.

Another object of the invention is an antibody containing at least one, at least two or all three HVR VL sequences selected from (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 122-124; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 125; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74. In one of the embodiments of the invention, the antibody contains (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 122-124; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 125; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74.

Another object of the invention is an antibody containing at least one, at least two or all three HVR VL sequences selected from (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 122; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (c) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 74. In one embodiment, the antibody contains (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 122; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (c) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 74. Another object of the invention is an antibody containing at least one, at least two, or all three HVR VL sequences selected from (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 123; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (c) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 74. In one embodiment, the antibody contains (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 123; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (c) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 74.

Another object of the invention is an antibody containing at least one, at least two or all three HVR VL sequences selected from (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 129; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 130; and (c) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 131. In one embodiment, the antibody contains (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 129; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 130; and (c) HVR-L3 which contains the amino acid sequence of SEQ ID NO 131.

According to another aspect of the invention, an antibody of the invention comprises (a) a VH domain comprising at least one, at least two, or all three HVR VH sequences selected from (I) HVR-H1, which comprises one of the amino acid sequences SEQ ID NOs: 55-57, 114, 115, 126, (II) HVR-H2, which contains one of the amino acid sequences of SEQ ID NOs: 58-60, 116-120, 127, and (III) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64, 121, 128; and (b) a VL domain comprising at least one, at least two, or all three HVR VL sequences selected from (I) HVR-L1, which comprises one of the amino acid sequences of SEQ ID NOs: 65-69, 122- 124, 129, (II) HVR-L2 which contains one of the amino acid sequences of SEQ ID NOs: 70-72, 125, 130, and (III) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74 , 131.

According to another aspect of the invention, an antibody of the invention comprises (a) a VH domain comprising at least one, at least two, or all three HVR VH sequences selected from (I) HVR-H1, which comprises one of the amino acid sequences SEQ ID NOs: 55-57, (II) HVR-H2 which contains one of the amino acid sequences of SEQ ID NOs: 58-60, and (III) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64 ; and (b) a VL domain containing at least one, at least two, or all three HVR VL sequences selected from (I) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 65-69, (II ) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72, and (III) HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73-74.

According to another aspect of the invention, an antibody of the invention comprises (a) a VH domain comprising at least one, at least two, or all three HVR VH sequences selected from (I) HVR-H1, which comprises one of the amino acid sequences SEQ ID NO: 114-115, (II) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 116-120, and (III) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 121; and (b) a VL domain containing at least one, at least two, or all three HVR VL sequences selected from (I) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 122-124, (II ) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 125, and (III) HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 73-74.

According to another aspect of the invention, an antibody of the invention comprises (a) a VH domain comprising at least one, at least two, or all three HVR VH sequences selected from (I) HVR-H1, which comprises the amino acid sequence of SEQ ID NO: 114, (II) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58, and (III) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63; and (b) a VL domain comprising at least one, at least two, or all three HVR VL sequences selected from (I) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 122, (II) HVR-L2 , which contains the amino acid sequence of SEQ ID NO: 71, and (III) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 74. According to another object of the invention, the antibody of the invention contains (a) a VH domain containing at least at least one, at least two or all three HVR VH sequences selected from (I) HVR-H1 which contains the amino acid sequence of SEQ ID NO: 114, (II) HVR-H2 which contains the amino acid sequence of SEQ ID NO: 58, and (III) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63; and (b) a VL domain comprising at least one, at least two, or all three HVR VL sequences selected from (I) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 123, (II) HVR-L2 , which contains the amino acid sequence of SEQ ID NO: 71, and (III) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 74.

According to another aspect of the invention, an antibody of the invention comprises (a) a VH domain comprising at least one, at least two, or all three HVR VH sequences selected from (I) HVR-H1, which comprises the amino acid sequence of SEQ ID NO: 126, (II) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 127, and (III) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 128; and (b) a VL domain comprising at least one, at least two, or all three HVR VL sequences selected from (I) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 129, (II) HVR-L2 , which contains the amino acid sequence of SEQ ID NO: 1301, and (III) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 131.

Another object of the invention is an antibody containing (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55-57, 114-115, 126; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60, 116-120, 127; (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64, 121, 128; (d) HVR-L1 which contains one of the amino acid sequences of SEQ ID NOs: 65-69, 122-124, 129; (e) HVR-L2 which contains one of the amino acid sequences of SEQ ID NOs: 70-72, 125, 130; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74, 131.

Another object of the invention is an antibody containing (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55-57, 114-115; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58-60, 116-120; (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 61-64, 121; (d) HVR-L1 which contains one of the amino acid sequences of SEQ ID NOs: 65-69, 122-124; (e) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72, 125; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74.

Another object of the invention is an antibody containing (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 114-115; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 116-120; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 121; (d) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 122-124; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 125; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74.

Another object of the invention is an antibody containing (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 114; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63; (d) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 122; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (e) HVR-L3, which contains the amino acid sequence of SEQ ID NO: 74. Another object of the invention is an antibody containing (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 114; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 63; (d) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 123; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (e) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 74.

Another object of the invention is an antibody containing (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 126; (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 127; (c) HVR-H3, which contains the amino acid sequence of SEQ ID NO: 128; (d) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 129; (e) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 130; and (e) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 131.

In some embodiments, any one or more amino acids in the anti-myostatin antibody provided herein are substituted at the following HV positions: (a) HVR-H1 (SEQ ID NO: 55) at positions 1 and 2; (b) in HVR-H2 (SEQ ID NO: 58) at positions 4, 7, 8, 10, 11, 12 and 16; (c) in HVR-H3 (SEQ ID NO: 61) at positions 5, 7 and 11; (d) in HVR-L1 (SEQ ID NO: 65) at positions 1, 2, 5, 7, 8 and 9; (e) in HVR-L2 (SEQ ID NO: 70) at positions 3 and 7; and (e) in HVR-L3 (SEQ ID NO: 73) at position 8.

In some embodiments of the invention, one or more amino acid(s) substitution(s) in the antibody to myostatin is(are) a conservative(s) substitution(s) presented in the present description. In some embodiments, one or more of the following amino acid substitutions may be made in any combination: (a) to HVR-H1 (SEQ ID NO: 55) S1H; Y2T, D or E; (b) in HVR-H2 (SEQ ID NO: 58) Y4H; S7K; T8M or K; Y10K; A11M or E; S12E; G16K; (c) in HVR-H3 (SEQ ID NO: 61) Y5H; T7H; L11K; (d) in HVR-L1 (SEQ ID NO: 65) Q1T; S2T; S5E; Y7F; D8H; N9D or A or E; (e) in HVR-L2 (SEQ ID NO: 70) S3E; S7Y, F or W; and (e) in HVR-L3 (SEQ ID NO: 73) L8R.

All possible combinations of the above substitutions are included in the consensus sequences of SEQ ID NOs: 126, 127, 128, 129, 130 and 131 HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2 and HVR-L3, respectively.

In any of the above embodiments, the anti-myostatin antibody may be humanized. In one embodiment, the anti-myostatin antibody comprises an HVR as defined in any of the above embodiments and further comprises an acceptor human framework, such as a human immunoglobulin framework or a human consensus framework. In another embodiment, the anti-myostatin antibody comprises an HVR as defined in any of the above embodiments and further comprises a VH or VL containing the FR sequence. In a further embodiment, the anti-myostatin antibody comprises the following FR sequences of the heavy chain and/or light chain variable domain: in the heavy chain variable domain, FR1 contains one of the amino acid sequences of SEQ ID NO: 132-134, FR2 contains one of the amino acid sequences of SEQ ID NO: : 135-136, FR3 contains one of the amino acid sequences of SEQ ID NO: 137, FR4 contains one of the amino acid sequences of SEQ ID NO: 138. In the light chain variable domain, FR1 contains the amino acid sequence of SEQ ID NO: 139, FR2 contains one of the amino acid sequences SEQ ID NO: 140-141, FR3 contains one of the amino acid sequences of SEQ ID NO: 142-143, FR4 contains the amino acid sequence of SEQ ID NO: 144.

One of the objects of the invention is an antibody to myostatin, containing at least one, two, three, four, five or six HVR selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 157-162; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 163-168; (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 169-174; (d) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 175-180; (e) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 181-186; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 187-192.

One of the objects of the invention is an antibody containing at least one, at least two or all three HVR VH sequences selected from (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 157-162; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 163-168; and (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 169-174. In one of the embodiments of the invention, the antibody contains HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 169-174. In another embodiment, the antibody comprises HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 169-174 and HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 187-192. In another embodiment, the antibody comprises HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 169-174, HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 187-192, and HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 163-168. In the following embodiment of the invention, the antibody contains (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 157-162; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 163-168; and (c) HVR-H3 which contains one of the amino acid sequences of SEQ ID NOs: 169-174.

Another object of the invention is an antibody containing at least one, at least two or all three HVR VL sequences selected from (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 175-180; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 181-186; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 187-192. In one of the embodiments of the invention, the antibody contains (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 175-180; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 181-186; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 187-192.

Another object of the invention is an antibody according to the invention, which contains (a) a VH domain containing at least one, at least two or all three HVR VH sequences selected from (I) HVR-H1, which contains one of the amino acids sequences of SEQ ID NO: 157-162, (II) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 163-168, and (III) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 169- 174; and (b) a VL domain containing at least one, at least two, or all three HVR VL sequences selected from (I) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 175-180, (II ) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 181-186, and (III) HVR-L3, which contains one of the amino acid sequences of SEQ ID NO: 187-192.

Another object of the invention is an antibody containing (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 157-162; (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 163-168; (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 169-174; (d) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 175-180; (e) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 181-186; and (e) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 187-192.

In another aspect of the invention, an anti-myostatin antibody comprises a heavy chain variable domain (VH) sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99 % or 100% identical to one of the amino acid sequences of SEQ ID NOs: 13, 16-30, 32-34 and 86-95. In some embodiments, a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical contains substitutions (e.g., conservative substitutions ), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 13, 16-30, and 32-34. In some embodiments, the substitutions, insertions, or deletions affect regions outside of the HVR (ie, FR). Optionally, the anti-myostatin antibody contains the VH sequence shown in one of SEQ ID NOs: 13, 16-30, and 32-34, including post-translational modifications of said sequence. In a particular embodiment, the VH contains one, two, or three HVRs selected from: (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NOs: 55-57, (b) HVR-H2, which contains one of the amino acids sequences of SEQ ID NO: 58-60, and (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64.

In another aspect of the invention, an anti-myostatin antibody comprises a heavy chain variable domain (VH) sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99 % or 100% identical to one of the amino acid sequences of SEQ ID NOs: 13, 16-30, 32-34 and 86-95. In some embodiments, a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical contains substitutions (e.g., conservative substitutions ), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 13, 16-30, 32-34, and 86-95. In some embodiments, the substitutions, insertions, or deletions affect regions outside of the HVR (ie, FR). Optionally, the anti-myostatin antibody contains the VH sequence shown in one of SEQ ID NOs: 13, 16-30, 32-34, and 86-95, including post-translational modifications of said sequence. In a particular embodiment, the VH contains one, two or three HVRs selected from: (a) HVR-H1 which contains one of the amino acid sequences of SEQ ID NOs: 55-57, 114-115, 126, (b) HVR-H2 , which contains one of the amino acid sequences of SEQ ID NOs: 58-60, 116-120, 127, and (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NOs: 61-64, 121, 128. Post-translational modifications include, but are not limited to, modification by pyroglutamilation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, an anti-myostatin antibody contains a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one from the amino acid sequences of SEQ ID NOs: 13, 16-30, 32, 33, and 34. In some embodiments, a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95% identical, 96%, 97%, 98%, or 99% contains substitutions (eg, conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-myostatin antibody containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 13, 16-30, 32, 33, and 34. In some embodiments, the substitution, insertion, or deletion affect areas outside the HVR (i.e. FR). Optionally, the anti-myostatin antibody contains the VH sequence shown in one of SEQ ID NOs: 13, 16-30, 32, 33 and 34, including post-translational modifications of said sequence. In a particular embodiment, the VH contains one, two, or three HVRs selected from: (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NOs: 55-57, (b) HVR-H2, which contains one of the amino acids sequences of SEQ ID NO: 58-60, and (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61-64. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, an anti-myostatin antibody contains a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one from the amino acid sequences of SEQ ID NOs: 86-95. In some embodiments, a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical contains substitutions (e.g., conservative substitutions ), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 86-95. In some embodiments, the substitutions, insertions, or deletions affect regions outside of the HVR (ie, FR). Optionally, the anti-myostatin antibody contains the VH sequence shown in one of SEQ ID NOs: 86-95, including post-translational modifications of said sequence. In a specific embodiment of the invention, the VH contains one, two or three HVRs selected from: (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 57, 114-115, 126, (b) HVR-H2, which contains one of the amino acid sequences of SEQ ID NO: 58, 116-120, 127, and (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 63, 121, 128. Post-translational modifications include (but not limited only) modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, the anti-myostatin antibody contains a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid identical. SEQ ID NO: 86. In some embodiments, a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, contains substitutions (eg, conservative substitutions), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in SEQ ID NO: 86. In some embodiments, the substitutions, insertions, or deletions affect regions outside the HVR (i.e., FR). Optionally, the anti-myostatin antibody contains the VH sequence shown in SEQ ID NO: 86, including post-translational modifications of said sequence. In a specific embodiment, the VH contains one, two, or three HVRs selected from: (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 114, (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58 , and (c) HVR-H3 which contains the amino acid sequence of SEQ ID NO: 63. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, the anti-myostatin antibody contains a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid identical. sequences of SEQ ID NO: 92. In some embodiments, a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, contains substitutions (eg, conservative substitutions), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in SEQ ID NO: 92. In some embodiments, the substitutions, insertions, or deletions affect regions outside the HVR (i.e., FR). Optionally, the anti-myostatin antibody contains the VH sequence shown in SEQ ID NO: 92, including post-translational modifications of said sequence. In a specific embodiment, the VH contains one, two, or three HVRs selected from: (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 114, (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 58 , and (c) HVR-H3 which contains the amino acid sequence of SEQ ID NO: 63. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, an anti-myostatin antibody contains a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one from the amino acid sequences of SEQ ID NOs: 15, 31, 35-38 and 96-99. In some embodiments, a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical contains substitutions (e.g., conservative substitutions ), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 15, 31, 35-38, and 96-99. In some embodiments, the substitutions, insertions, or deletions affect regions outside of the HVR (ie, FR). Optionally, the anti-myostatin antibody contains the VL sequence shown in one of SEQ ID NOs: 15, 31, 35-38 and 96-99, including post-translational modifications of said sequence. In a particular embodiment, the VL contains one, two or three HVRs selected from: (a) HVR-L1 which contains one of the amino acid sequences of SEQ ID NOs: 65-69, 122-124, 129; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72, 125, 130; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74, 131. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, an anti-myostatin antibody contains a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one from the amino acid sequences of SEQ ID NOs: 15, 31, 35, 36, 37, and 38. In some embodiments, a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95% identical, 96%, 97%, 98%, or 99% contains substitutions (eg, conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-myostatin antibody containing the indicated sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 15, 31, 35, 36, 37, and 38. In some embodiments, the substitution, insertion, or deletion affect areas outside the HVR (i.e. FR). Optionally, the anti-myostatin antibody contains the VL sequence shown in one of SEQ ID NOs: 15, 31, 35, 36, 37 and 38, including post-translational modifications of said sequence. In a specific embodiment, the VL contains one, two or three HVRs selected from: (a) HVR-L1 which contains one of the amino acid sequences of SEQ ID NOs: 65-69; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70-72; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NOs: 73-74. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, an anti-myostatin antibody contains a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one from the amino acid sequences of SEQ ID NOs: 96-99. In some embodiments, a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical contains substitutions (e.g., conservative substitutions ), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 96-99. In some embodiments, the substitutions, insertions, or deletions affect regions outside of the HVR (ie, FR). Optionally, the anti-myostatin antibody contains the VL sequence shown in one of SEQ ID NOs: 96-99, including post-translational modifications of said sequence. In a specific embodiment, the VL contains one, two or three HVRs selected from: (a) HVR-L1 which contains one of the amino acid sequences of SEQ ID NOs: 122-124, 129; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 71, 125, 130; and (c) HVR-L3, which contains the amino acid sequence of SEQ ID NOs: 74, 131. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain in pyroglutamic acid.

In another aspect of the invention, the anti-myostatin antibody contains a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid identical. SEQ ID NO: 96. In some embodiments, a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, contains substitutions (eg, conservative substitutions), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in SEQ ID NO: 96. In some embodiments, the substitutions, insertions, or deletions affect regions outside the HVR (i.e., FR). Optionally, the anti-myostatin antibody contains the VL sequence shown in SEQ ID NO: 96, including post-translational modifications of said sequence. In a specific embodiment of the invention, the VL contains one, two or three HVRs selected from: (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 122; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (c) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 74. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, the anti-myostatin antibody contains a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid identical. sequences of SEQ ID NO: 97. In some embodiments, a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical, contains substitutions (eg, conservative substitutions), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in SEQ ID NO: 97. In some embodiments, the substitutions, insertions, or deletions affect regions outside the HVR (i.e., FR). Optionally, the anti-myostatin antibody contains the VL sequence shown in SEQ ID NO: 97, including post-translational modifications of said sequence. In a specific embodiment of the invention, the VL contains one, two or three HVRs selected from: (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 123; (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 71; and (c) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 74. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

Another object of the invention is an antibody to myostatin, where the antibody contains VH, presented in one of the above embodiments of the invention, and VL, presented in one of the above embodiments of the invention. In one embodiment, the antibody comprises the VH and VL sequences shown in one of SEQ ID NOs: 13, 16-30, 32-34, and 86-95 and one of SEQ ID NOs: 15, 31, 35-38 and 96-99, respectively, including post-translational modifications of these sequences. In one of the embodiments of the invention, the antibody contains the VH and VL sequences indicated in one of SEQ ID NOs: 13, 16-30 and 32-34 and in one of SEQ ID NOs: 15, 31 and 35-38, respectively, including post- translational modifications of these sequences. In one embodiment, the antibody comprises the VH and VL sequences shown in one of SEQ ID NOs: 86-95 and one of SEQ ID NOs: 96-99, respectively, including post-translational modifications of these sequences. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

Another object of the invention is an antibody to myostatin, where the antibody contains VH, presented in any of the above embodiments of the invention, and VL, presented in any of the above embodiments of the invention. In one embodiment, the antibody comprises the VH and VL sequences of SEQ ID NO: 86 and SEQ ID NO: 96, respectively, including post-translational modifications of these sequences. In one embodiment, the antibody comprises the VH and VL sequences of SEQ ID NO: 92 and SEQ ID NO: 97, respectively, including post-translational modifications of these sequences. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

Another object of the invention is an antibody to myostatin containing a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one from the amino acid sequences of SEQ ID NO: 12, 145-150. In some embodiments, a VH sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical contains substitutions (e.g., conservative substitutions ), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted and/or deleted by deletion in one of SEQ ID NOs: 12, 145-150. In some embodiments, the substitutions, insertions, or deletions affect regions outside of the HVR (ie, FR). Optionally, the anti-myostatin antibody contains the VH sequence shown in one of SEQ ID NOs: 12, 145-150, including post-translational modifications of said sequence. In a particular embodiment, the VH contains one, two, or three HVRs selected from: (a) HVR-H1, which contains one of the amino acid sequences of SEQ ID NO: 55, 157-162, (b) HVR-H2, which contains one from the amino acid sequences of SEQ ID NO: 58, 163-168, and (c) HVR-H3, which contains one of the amino acid sequences of SEQ ID NO: 61, 169-174. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In another aspect of the invention, an anti-myostatin antibody contains a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one from the amino acid sequences of SEQ ID NO: 14, 151-156. In some embodiments, a VL sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical contains substitutions (e.g., conservative substitutions ), insertions or deletions relative to the reference sequence, but an antibody to myostatin containing the specified sequence retains the ability to bind to myostatin. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in one of SEQ ID NOs: 14, 151-156. In some embodiments, the substitutions, insertions, or deletions affect regions outside of the HVR (ie, FR). Optionally, the anti-myostatin antibody contains the VL sequence shown in one of SEQ ID NOs: 14, 151-156, including post-translational modifications of said sequence. In a specific embodiment of the invention, the VL contains one, two or three HVRs selected from: (a) HVR-L1, which contains one of the amino acid sequences of SEQ ID NO: 65, 175-180; (b) HVR-L2, which contains one of the amino acid sequences of SEQ ID NO: 70, 181-186; and (c) HVR-L3 which contains one of the amino acid sequences of SEQ ID NO: 73, 187-192. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

Another object of the invention is an antibody to myostatin, where the antibody contains VH, presented in any of the above embodiments of the invention, and VL, presented in any of the above embodiments of the invention. In one embodiment, the antibody comprises the VH and VL sequences shown in one of SEQ ID NOs: 12, 145-150 and one of SEQ ID NOs: 14, 151-156, respectively, including post-translational modifications of these sequences. Post-translational modifications include, but are not limited to, modification by pyroglutamylation of glutamine or glutamate at the N-terminal region of the heavy chain or light chain to pyroglutamic acid.

In some embodiments, an anti-myostatin antibody of the present invention comprises a VH as shown in any of the above embodiments and a heavy chain constant region comprising one of the amino acid sequences of SEQ ID NOs: 7, 9, 11, 193, 195-198, 227, 228, 229-381. In some embodiments, an anti-myostatin antibody of the present invention comprises a VL as defined in any of the above embodiments and a light chain constant region comprising one of the amino acid sequences of SEQ ID NOs: 8 and 10.

The following object of the invention is an antibody that binds to the same epitope as the antibody to myostatin presented in the present description. The next object of the invention is an antibody that binds to the same epitope as the antibody listed in table 2A. Another aspect of the invention is an antibody that binds to the same epitope as the antibody listed in Tables 11a or 13. In some embodiments, the invention is an antibody that binds to an epitope within a myostatin propeptide fragment consisting of amino acids 21-100 of SEQ ID NO: 78. Alternatively, the antibody binds to a fragment of the myostatin propeptide, which consists of amino acids 21-80, 41-100, 21-60, 41-80, 61-100, 21-40, 41-60, 61-80 or 81- 100 SEQ ID NO: 78.

In another embodiment, the anti-myostatin antibody of any of the above embodiments is a monoclonal antibody, including a chimeric, humanized, or human antibody. In one embodiment, the anti-myostatin antibody is an antibody fragment, such as an Fv, Fab, Fab', dimeric antibody, or F(ab') ₂ fragment. In another embodiment, the antibody is a full-length antibody, for example, an intact antibody of an IgG1 or IgG4 isotype, or another class or isotype of antibodies described herein.

In a further aspect of the invention, the anti-myostatin antibody referred to in any of the above embodiments of the invention may have any of the features, singly or in combination, as described in sections 1-7 below:

1. Antibody affinity

In some embodiments, an antibody provided herein has a dissociation constant (Kd) value of 1 μM or less, 100 nM or less, 10 nM or less, 1 nM or less, 0.1 nM or less, 0, 0 1 nM or less, or 0.001 nM or less (for example, 10 ^-8 M or less, for example, from 10 ^-8 M to 10 ^-13 M, for example, from 10 ^-9 M to 10 ^-13 M).

In one embodiment, the Kd value is measured using a radiolabeled antigen (RIA) binding assay. In one of the embodiments of the invention, RIA is carried out using the Fab version of the antibody of interest and its antigen. For example, the binding affinity of a Fab to an antigen is measured in solution by equilibrating the Fab with a minimum concentration of ¹²⁵ I-labeled antigen in the presence of titrated dilutions of unlabeled antigen, then "capturing" the bound antigen on an anti-Fab sensitized plate (see, e.g., Chen et al. ., J. Mol Biol 293, 1999, pp. 865-881). ^MICROTITER® multiwell plates (Thermo Scientific) are sensitized overnight with anti-Fab capture antibody (Cappel Labs) at 5 µg/mL in 50 mM sodium carbonate (pH 9.6) and then blocked with 2% (w/v) bovine serum albumin in PBS for 2-5 hours at room temperature (about 23°C). In a non-absorbent plate (Nunc, No. 269620), 100 or 26 pM ¹²⁵ I-labeled antigen is mixed with serial dilutions of the Fab of interest (e.g., anti-VEGF antibody, Fab-12, suitable for evaluating, see Presta et al. , Cancer Res. 57, 1997, pp. 4593-4599). The Fab of interest is then incubated overnight; however, the incubation can be carried out for a longer period of time (for example, for 65 hours) in order to ensure that equilibrium is reached. The mixtures are then transferred to a prepared capture plate and incubated at room temperature (eg, for 1 hour). The solution is then removed and the plate washed eight times with 0.1% polysorbate 20 (TWEEN- ^20® ) in PBS. After the plates are dry, 150 µl/well of scintillation fluid ((MICROSCINT-20™; Packard) is added to the plates and the plates are read using a TOPCOUNT™ gamma counter (Packard) for 10 minutes. For competitive binding assays choose the concentration of each Fab, providing a level of binding that is less than or equal to 20% of the maximum.

In another embodiment of the invention, Kd is measured by surface plasmon resonance using a BIACORE® device. For example, the assay is performed with a ^BIACORE® -2000 or ^BIACORE® -3000 device (GE Healthcare Inc., Piscataway, NJ) at 25° C. using antigen immobilized on CM5 chips with an immobilization level corresponding to ~ 10 response units (RU). In one embodiment, carboxymethylated dextran (CM5, GE Healthcare Inc.) biosensor chips are activated with N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) hydrochloride according to the supplier's instructions. . The antigen is diluted with 10 mM sodium acetate, pH 4.8 to a concentration of 5 μg/ml (~0.2 μM) before injection at a flow rate of 5 μl/min to achieve an immobilization level corresponding to about 10 response units (RU) of the cross-linked protein. After injection of the antigen, 1 M ethanolamine is injected to block the unreacted groups. For kinetic measurements, two-fold serial dilutions of Fab (from 0.78 to 500 nM) in PBS supplemented with 0.05% polysorbate 20 (TWEEN-20™) as a surfactant (PBS) are injected at 25°C with a flow rate of approximately 25 μl /min The rates of the association reaction (k _on ) and the dissociation reaction (k _off ) are calculated using a simple 1:1 Langmuir binding model (Evaluation software version 3.2, BIACORE®) by simultaneously fitting the association and dissociation sensorgrams. The equilibrium constant of the dissociation reaction (Kd) is calculated as the ratio k _off /k _on (see, for example, Chen et al., J Mol Biol 293, 1999, pp. 865-881). If the association reaction rate exceeds 10 ⁶ M ^-1 s ^-1 as assessed by the surface plasmon resonance method described above, then the association reaction rate can be determined using the fluorescence quenching method, which measures the increase or decrease in the intensity of fluorescence emission (excitation wavelength 295 nm; emission wavelength 340 nm, bandwidth 16 nm) at 25°C applied at a concentration of 20 nM anti-antigen antibody (Fab-format) in PBS, pH 7.2, in the presence of increasing concentrations of antigen, by measuring with a spectrometer such as as a spectrophotometer equipped with a flow stop unit (Aviv Instruments) or an SLM-AMINCO™ 8000 series spectrophotometer (ThermoSpectronic) while stirring the contents of the cuvette.

2. Antibody fragments

In some embodiments, the antibody of the present invention is an antibody fragment. Antibody fragments include, but are not limited to, Fab, Fab', Fab'-SH, F(ab') ₂ , Fv, and scFv fragments, and other fragments described below. For a review of some antibody fragments, see Hudson et al., Nat. Med. 9, 2003, pp. 129-134. For a review of scFv fragments, see, for example, Plueckthun in: The Pharmacology of Monoclonal Antibodies, ed. Rosenburg and Moore, Springer, New York, Vol. 113, 1994, pp. 269-315; see also WO 93/16185; and US Nos. 5,571,894 and 5,587,458. For a discussion of Fab and F(ab') ₂ fragments that contain rescue receptor-binding epitope residues and have an extended in vivo half-life, see US No. 5,869,046.

Dimeric antibodies are antibody fragments with two antigen-binding sites, which can be divalent or bispecific (see, for example, EP 0404097; WO 1993/01161; Hudson et al., Nat. Med. 9, 2003, pp. 129-134; and Holliger et al., Proc Natl Acad Sci USA 90, 1993, pp. 6444-6448). Trimeric and tetrameric antibodies are also described in Hudson et al., Nat. Med. 9, 2003, pp. 129-134).

Single domain antibodies are antibody fragments containing all or part of the heavy chain variable domain, or all or part of the light chain variable domain of an antibody. In some embodiments, the single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, MA; see, for example, US No. 6248516 B1).

Antibody fragments can be generated by a variety of techniques including, but not limited to, proteolytic cleavage of an intact antibody, and also generated using recombinant host cells (eg, E. coli or phage) as described herein.

3. Chimeric and humanized antibodies

In some embodiments of the invention, the antibody specified in the present description is a chimeric antibody. Some chimeric antibodies are described, for example, in US No. 4816567; and Morrison et al., Proc. Natl. Acad. sci. USA 81, 1984, pp. 6851-6855. In one example, the chimeric antibody comprises a non-human variable region (eg, a variable region from an antibody from mice, rats, hamsters, rabbits, or non-human primates such as monkeys) and a human constant region. In another example, the chimeric antibody is a "class-switched" antibody whose class or subclass has been changed from the parent antibody. Chimeric antibodies include their antigen-binding fragments.

In some embodiments, the chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity in humans while retaining the specificity and affinity of the parent non-human antibody. Typically, a humanized antibody contains one or more variable domains in which the HVRs, eg CDRs (or portions thereof), are derived from a non-human antibody and the FRs (or portions thereof) are derived from human antibody sequences. The humanized antibody may optionally contain at least a portion of a human constant region. In some embodiments, certain FR residues in a humanized antibody are replaced with corresponding residues from a non-human antibody (eg, an antibody from which HVR residues are derived), for example, to maintain or improve the specificity or affinity of the antibody.

Information about humanized antibodies and methods for their creation are summarized, for example, in Almagro, Front. biosci. 13, 2008, pp. 1619-1633, and are also described in Riechmann et al., Nature 332, 1988, pp. 323-329; Queen et al., Proc. Natl. Acad. sci. USA 86, 1989, pp. 10029-10033; US Nos. 5821337, 7527791, 6982321 and 7087409; Kashmiri et al., Methods 36, 2005, pp. 25-34 (description of transplantation of SDR (a-CDR)); Padlan, Mol. Immunol. 28, 1991, pp. 489-498 (description of "recoating"); Dall'Acqua et al., Methods 36, 2005, pp. 43-60 (description of "FR permutation"); and Osbourn et al., Methods 36, 2005, pp. 61-68 and Klimka et al., Br. J. Cancer 83, 2000, pp. 252-260 (description of the "directed selection" approach for FR permutation).

Human framework regions that can be used for humanization include, but are not limited to: framework regions selected using "best fit" methods (see, for example, Sims et al., J. Immunol. 151, 1993, pp. 2296-2308, framework regions derived from the consensus sequence of a specific subset of human antibody light and heavy chain variable regions (see, e.g., Carter et al., Proc. Natl. Acad. Sci. USA 89, 1992, pp. 4285 -4289 and Presta et al., J. Immunol. 151, 1993, pp. 2623-2632), human mature (with somatic mutations) or human germline frameworks (see, for example, Almagro and Fransson, Front. Biosci. 13, 2008, pp. 1619-1633), and framework regions resulting from screening of FR libraries (see, e.g., Vasa et al., J. Biol. Chem. 272, 1997, pp. 10678-10684 and Rosok et al., J Biol Chem 271 (1996) 22611-22618).

4. Human antibodies

In some embodiments, the antibody of the present invention is a human antibody. Human antibodies can be generated using a variety of techniques known in the art. Human antibodies are described in general by van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5, 2001, pp. 368-374 and Lonberg, Curr. Opin. Immunol. 20, 2008, pp. 450-459.

Human antibodies can be generated by administering an immunogen to a transgenic animal modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigen challenge. These animals typically contain all or part of the human immunoglobulin loci instead of endogenous immunoglobulin loci, which are either present outside the chromosomes or integrated arbitrarily into the chromosomes of the animal. In such transgenic mice, endogenous immunoglobulin loci are usually inactivated. For a review of methods for generating human antibodies in transgenic animals, see Lonberg, Nat. Biotech. 23, 2005, pp. 1117-1125 (see, for example, also US No. 6075181 and US No. 6150584 for the XENOMOUSE™ Technology; US No. 5770429 for the HUMAB® Technology; US No. 7041870 for the K-M MOUSE® Technology and US Patent Application Publication 2007/0061900 description of VELOCIMOUSE® technology). Human variable regions from intact antibodies obtained from these animals can be further modified, for example by combining with another human constant region.

Human antibodies can be generated using hybridoma-based methods. Human myeloma and murine-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described (see, for example, Kozbor, J. Immunol. 133, 1984, cc. 3001-3005; Brodeur et al., Monoclonal Antibody Production Techniques and Applications , Marcel Dekker, Inc., New York, 1987, pp. 51-63 and Boerner et al., J. Immunol. 147, 1991, pp. 86-95). Human antibodies generated using human B-cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. sci. USA 103, 2006, pp. 3557-3562. Additional methods include those described, for example, in US No. 7189826 (description of obtaining monoclonal human antibodies of the IgM type from hybridoma cell lines), and Ni, Xiandai Mianyixue 26, 2006, cc. 265-268 (description of human-human hybridomas). Human hybridoma technology (Trioma technology) is also described by Vollmers and Brandlein, Histology and Histopathology 20, 2005, cc. 927-937 and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology 27, 2005, pp. 185-191.

Human antibodies can also be generated by isolating the variable domain sequences of Fv clones selected from phage display libraries that have been generated using human antibodies. These variable domain sequences can then be combined with the desired human constant domain. Methods for selecting human antibodies from antibody libraries are described below.

5. Antibodies obtained from libraries

Antibodies of the invention can be isolated by screening combinatorial libraries of antibodies with the desired activity or activities. For example, a variety of methods are known in the art for generating phage display libraries and screening said libraries for antibodies having the desired binding characteristics. These methods are summarized in, for example, Hoogenboom et al., Methods in Molecular Biology 178, 2001, cc. 1-37 and are also described, for example, in McCafferty et al., Nature 348, 1990, cc. 552-554; Clackson et al., Nature 352, 1991, pp. 624-628; Marks et al., J. Mol. Biol. 222, 1992, pp. 581-597; Marks and Bradbury, Methods in Molecular Biology 248, 2003, pp. 161-175; Sidhu et al., J. Mol. Biol. 338, 2004, pp. 299-310; Lee et al., J. Mol. Biol. 340, 2004, pp. 1073-1093; Fellouse, Proc. Natl. Acad. sci. USA 101, 2004, pp. 12467-12472 and Lee et al., J. Immunol. Methods 284, 2004, pp. 119-132.

In some phage display techniques, populations of VH and VL genes are cloned individually by polymerase chain reaction (PCR) and recombined randomly in phage libraries, which are then screened for antigen-binding phage according to the method described by Winter et al., Ann . Rev. Immunol. 12, 1994, pp. 433-455. The phage typically displays antibody fragments either as single chain Fv fragments (scFv) or as Fab fragments. Libraries derived from immunized sources include antibodies with high affinity for the immunogen without the need for hybridomas. Alternatively, an untreated ("naive") population (e.g. from a human body) can be cloned, producing a single source of antibodies to a wide range of foreign as well as self antigens without any immunization, as described in Griffiths et al., EMBO J, 12, 1993, pp. 725-734. Finally, "naive" libraries can also be obtained by synthetic methods by cloning non-rearranged V-gene segments from stem cells and using PCR primers containing a random sequence to encode hypervariable CDR3 regions and to perform in vitro rearrangement according to the method described in Hoogenboom and Winter, J. Mol. Biol. 227, 1992, pp. 381-388. Phage libraries of human antibodies are described, for example, in the following patent publications: US No. 5750373 and US Patent Application Publications 2005/0079574, 2005/0119455, 2005/0266000, 2007/0117126, 2007/0160598, 2007/0237764, 2007/0292936 and 2009/0002360.

Antibodies or antibody fragments isolated from human antibody libraries are considered to be human antibodies or human antibody fragments.

6. Multispecific antibodies

In some embodiments of the invention, the antibody specified in the present description is a multispecific antibody, for example, bispecific antibody. Multispecific antibodies are monoclonal antibodies that have specificities that bind to at least two different sites. In some embodiments, one of the binding specificities allows binding to myostatin and the other to any other antigen. In some embodiments, the bispecific antibodies can bind to two different myostatin epitopes. Bispecific antibodies can also be used to localize cytotoxic agents in cells that express myostatin. Bispecific antibodies can be obtained as full length antibodies or antibody fragments.

Techniques for generating multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (see Milstein and Cuello, Nature 305, 1983, pp. 537-540, WO 93/08829 and Traunecker et al., EMBO J. 10,1991, pp. 3655-3659), and "knob-in-hole" technology . Multispecific antibodies can also be obtained by creating controlled electrostatic influences to obtain antibody molecules with heterodimeric Fc (WO 2009/089004); cross-linking two or more antibodies or fragments (see, for example, US No. 4676980 and Brennan and others, Science 229, 1985, pp. 81-83); the use of leucine zippers to produce bispecific antibodies (see, for example, Kostelny et al., J. Immunol. 148, 1992, pp. 1547-1553); the use of "dimeric antibodies (diabodies)" technology to generate bispecific antibody fragments (see, for example, Holliger et al., Proc. Natl. Acad. Sci. USA 90, 1993, pp. 6444-6448); and the use of single chain Fv (sFv) dimers (see, for example, Gruber et al., J. Immunol. 152, 1994, pp. 5368-5374); and obtaining trispecific antibodies according to the method described, for example, Tutt and others, J. Immunol. 147, 1991, pp. 60-69).

Also within the scope of the present invention are engineered antibodies with three or more functional antigen-binding sites, including "octopus antibodies" (see, for example, US 2006/0025576 A1).

The antibody or fragment thereof may also include a "dual acting Fab" or "DAF" containing an antigen-binding site that binds to myostatin as well as to another antigen other than it (see, for example, US 2008/0069820).

7. Antibody variants

Some embodiments of the invention relate to variants of the amino acid sequences of the antibodies presented in the present description. For example, it may be desirable to increase the binding affinity and/or other biological properties of an antibody. Variants of the amino acid sequence of an antibody can be obtained by introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by synthesizing peptides. These modifications include, for example, deletions and/or insertions and/or substitutions of residues in the amino acid sequences of the antibody. Any combination of deletions, insertions, and substitutions can be used to produce the final construct, as long as the final construct has the desired characteristics, such as the ability to bind to an antigen.

a) Variants obtained by substitution, insertion and deletion

Some embodiments of the invention are antibody variants that have one or more amino acid substitutions. Sites of interest for displacement mutagenesis include HVR and FR. Conservative substitutions are listed below in Table 1 under the heading "preferred substitutions" and are described below with reference to the amino acid side chain classes. More important substitutions are presented below in Table 1 under the heading "exemplary substitutions" and are described below with reference to the classes of amino acid side chains. Amino acid substitutions can be introduced into the antibody of interest and the products screened for desired activity, eg, retention/increase in antigen binding, reduced immunogenicity, or increased ADCC or CDC.

Amino acids can be grouped based on common side chain properties as follows: (1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) main: His, Lys, Arg; (5) residues that affect chain orientation: Gly, Pro; (6) aromatic: Trp, Tyr, Phe. Non-conservative substitutions mean the replacement of a representative of one of the indicated classes with a representative from another class.

One type of substitution-derived variant involves substitution of one or more hypervariable region residues of the parent antibody (eg, humanized or human antibody). In general, the resulting variant(s) selected for further study should have modifications (e.g., improvements) in certain biological properties (e.g., increased affinity, reduced immunogenicity) relative to the parent antibody, and /or must (s) have practically retained certain biological properties of the parent antibody. An example of a substitution-derived variant is an affinity matured antibody, which can be generated, for example, using phage display-based affinity maturation technologies, such as those described herein. In general, the method is as follows: one or more HVR residues are mutated and antibody variants are displayed on phage and screened for a particular biological activity (eg, binding affinity).

Changes (eg substitutions) can be made in the HVR, eg to increase the affinity of the antibody. These changes can be made in "hot (active) spots" in the HVR, i.e. residues encoded by codons that mutate at a high rate during somatic maturation (see, for example, Chowdhury, Methods Mol. Biol. 207, 2008, pp. 179-196), and/or in residues that come into contact with antigen, followed by testing the VH or VL variant for binding affinity. Affinity maturation by building and reselecting from secondary libraries is described, for example, in Hoogenboom et al., Methods in Molecular Biology 178, 2002, cc. 1-37. In some embodiments, the affinity maturation process introduces diversity into the variable region genes selected for maturation using any of a wide variety of methods (eg, reduced precision PCR, strand shuffling, or site-directed mutagenesis using oligonucleotides). Then a secondary library is created. The library is then screened to identify any antibody variants with the desired affinity. Another method used to introduce diversity includes HVR-targeting approaches that randomize multiple HVR residues (eg, 4-6 residues at a time). HVR residues involved in antigen binding can be specifically identified, for example, using alanine-scanning mutagenesis or modeling. Often the targets are primarily CDR-H3 and CDR-L3.

In some embodiments, the substitutions, insertions, or deletions may affect one or more HVRs, as long as the changes do not significantly reduce the ability of the antibody to bind to the antigen. For example, conservative changes can be made to the HVR (eg conservative substitutions as described herein) that do not significantly reduce binding affinity. These changes, for example, may be outside of the HVR residues involved in contacting the antigen. In some embodiments, in the sequences of the VH and VL variants above, each HVR is either unchanged or contains no more than one, two, or three amino acid substitutions.

A valuable method for identifying antibody residues or regions that can be mutated is the so-called "alanine-scanning mutagenesis" described in Cunningham and Wells, Science 244, 1989, pp. 1081-1085. In this method, a residue or group of target residues (e.g., charged residues such as arg, asp, his, lys, and glu) are identified and replaced with a neutral or negatively charged amino acid (e.g., alanine or polyalanine) to decide whether whether this will affect the interaction of the antibody with the antigen. Additional substitutions can be introduced at amino acid positions that have been shown to be functionally sensitive to the initial substitutions. Alternatively or additionally, the crystal structure of the antigen-antibody complex can be analyzed to identify points of contact between the antibody and antigen. These contacting residues and neighboring residues can be considered as targets or excluded from consideration as candidates for replacement. Variants can be screened to determine if they have the desired properties.

Insertions into amino acid sequences include amino- and/or carboxy-terminal fusions ranging in length from one residue to polypeptides containing one hundred or more residues, as well as insertions of one or more amino acid residues into the sequence. An example of the result of a terminal insertion is an antibody with an N-terminal methionyl residue. Other insertion variants of antibody molecules include the fusion of the N- or C-terminus of the antibody with an enzyme (eg, for ADEPT (antibody-directed enzyme prodrug therapy) or a polypeptide that prolongs the serum half-life of the antibody.

b) Glycosylation variants

In some embodiments of the invention, the antibody of the invention is modified to increase or decrease the degree of glycosylation of the antibody. The addition or deletion of glycosylation sites in an antibody can be easily accomplished by changing the amino acid sequence in such a way that one or more glycosylation sites are created or removed.

If the antibody contains an Fc region, then you can change the carbohydrate attached to it. Native antibodies that are produced by mammalian cells typically contain a branched biantenna oligosaccharide that is typically N-linked to the Asn297 CH2 domain of the Fc region (see, for example, Wright et al., TIBTECH 15, 1997 , pp. 26-32). The oligosaccharide may include various carbohydrates, such as mannose, N-acetylglucosamine (GlcNAc), galactose and sialic acid, as well as fucose attached to GlcNAc in the stem of the biantennary oligosaccharide structure. In some embodiments, modifications to the oligosaccharide in an antibody of the invention can be made to create antibody variants with certain improved properties.

One of the embodiments of the invention are variants of the antibody having a carbohydrate structure in which the content of fucose attached (directly or indirectly) to the Fc region is reduced. For example, the fucose content in said Fc region may be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The fucose content is determined by calculating the average sugar chain fucose content per Asn297 relative to the sum of all glycostructures attached to Asn297 (e.g. complex, hybrid and high mannose structures), which is measured by MALDI-TOF mass spectrometry according to the method described, for example, in WO 2008/077546. Asn297 denotes an asparagine residue present at approximately position 297 in the Fc region (EU numbering of Fc region residues); however, due to minor sequence variations, the Asn297 antibody can also be located at a position about +3 amino acids upstream or downstream from position 297, i.e. between positions 294 and 300. These fucosylated variants may have improved ADCC function (see, for example, US 2003/0157108 (Presta L.); US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd.). "Defucosylated" or "fucose deficient" antibody variants are: US 2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614; US 2002/0164328; US 2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865; WO 2003/085119; WO 2003/084570; WO 2005/035586; WO 2005/035778; WO 200aki5/053742; WO 2002/053742; J. Mol. Biol. 336, 2004, pp. 1239-1249; Yamane-Ohnuki et al., Biotech. Bioeng. 87, 2004, pp. 614-622). Examples of cell lines that have the ability to produce defucosylated antibodies are protein fucosylation deficient Lec13 CHO cells (Ripka et al., Arch. Biochem. Biophys. 249, 1986, pp. 533-545; US 2003/0157108 (Presta L. ) and WO 2004/056312 (Adams et al., see especially Example 11), and gene-knocked cell lines, e.g. (see, for example, Yamane-Ohnuki et al., Biotech. Bioeng. 87, 2004, pp. 614-622); Kanda et al., Biotechnol. Bioeng., 94(4), 2006, pp. 680-688 ; and WO 2003/085107).

In addition, variants of antibodies with bisected oligosaccharides are described, for example, in which the bisected oligosaccharide attached to the Fc region of the antibody is bisected with GlcNAc. These antibody variants may have reduced fucosylation and/or increased ADCC function. Examples of these antibody variants are described, for example, in WO 2003/011878 (Jean-Mairet et al.); US No. 6602684 (Umana et al.) and US 2005/0123546 (Umana et al.). Antibody variants with at least one galactose residue in the oligosaccharide attached to the Fc region are also proposed. These antibody variants may have increased CDC function. These antibody variants are described, for example, in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju S.); and WO 1999/22764 (Raju S.).

c) Fc region variants

According to specific embodiments of the invention, one or more amino acid modifications can be introduced into the Fc region of an antibody provided herein, thereby creating a variant Fc region. A variant Fc region may comprise a human Fc region sequence (eg, human IgG1, IgG2, IgG3, or IgG4 Fc regions) comprising an amino acid modification (eg, substitution) at one or more amino acid positions.

Acad. Sci. USA 82, 1985, cc. 1499-1502); US №5821337 (см. Bruggemann и др., J. Exp. Med. 166, 1987, cc. 1351-1361). В альтернативном варианте можно применять нерадиоактивные методы анализа (см., например, нерадиоактивный анализ цитотоксичности АСТ1™ на основе проточной цитометрии (фирма CellTechnology, Inc. Маунтин-Вью, шт. Калифорния); и нерадиоактивный анализ цитотоксичности CytoTox 96^® (фирма Promega, Мэдисон, шт. Висконсин). Пригодными для таких анализов эффекторными клетками являются мононуклеарные клетки периферической крови (РВМС) и естественные клетки-киллеры (NK). В альтернативном или дополнительном варианте ADCC-активность представляющей интерес молекулы можно оценивать in vivo, например, на животной модели, например, как описано y Clynes и др., Proc. Natl. Acad. Sci. USA 95, 1998, cc. 652-656. Можно осуществлять также анализы связывания C1q для подтверждения того, что антитело не может связывать C1q и поэтому у него отсутствует CDC-активность (см., например, описание ELISA для оценки связывания C1q и С3с в WO 2006/029879 и WO 2005/100402). Для оценки активации комплемента можно осуществлять анализ CDC (см., например, Gazzano-Santoro и др., J. Immunol. Methods 202, 1996, сс.163-171; Cragg и др., Blood 101, 2003, сс. 1045-1052; и Cragg, Blood 103, 2004, сс. 2738-2743). Определение FcRn-связывания и клиренса/времени полужизни in vivo можно осуществлять также с использованием методов, известных в данной области (см., например, Petkova и др.,

. Immunol. 18(12), 2006, сс.1759-1769).Some embodiments of the invention relate to an antibody variant that has some but not all effector functions, making it a promising candidate for applications where the in vivo half-life of the antibody is important, but some effector functions (such as CDC and ADCC) are not are necessary or harmful. In vitro and/or in vivo cytotoxicity assays can be performed to confirm the reduction/depletion of CDC and/or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks the ability to bind to Fc-gamma R (and therefore likely lacks ADCC activity), but retains the ability to bind to FcRn. Primary cells mediating ADCC, ie. NK cells express only Fc-gamma RIII while monocytes express Fc-gamma RI, Fc-gamma RII and Fc-gamma RIII. FcR expression data on hematopoietic cells are summarized in Table 3 on p. 464 by Ravetch and Kinet, Annu. Rev. Immunol. 9, 1991, p. 457-492. Examples of in vitro assays for ADCC activity of a molecule of interest are (but not limited to) those described in US No. 5500362 (see, for example, Hellstrom et al., Proc. Natl. Acad. Sci. USA 83, 1986, pp. 7059-7063 and Hellstrom et al., Proc.

Acad. sci. USA 82, 1985, pp. 1499-1502); US No. 5821337 (see Bruggemann et al., J. Exp. Med. 166, 1987, cc. 1351-1361). Alternatively, non-radioactive assays can be used (see, for example, non-radioactive flow cytometry-based ACT1™ cytotoxicity assay (CellTechnology, Inc. Mountain View, CA); and non-radioactive CytoTox ^96® cytotoxicity assay (Promega, Madison). , Wisconsin.) Suitable effector cells for such assays are peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively, or in addition, the ADCC activity of a molecule of interest can be assessed in vivo, for example, in an animal model , for example, as described by Clynes et al., Proc. Natl. Acad. Sci. USA 95, 1998, pp. 652-656 C1q binding assays can also be performed to confirm that the antibody cannot bind C1q and therefore no CDC activity (see, for example, WO 2006/029879 and WO 2005/100402 ELISA for assessing C1q and C3c binding). to analyze the CDC (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202, 1996, pp. 163-171; Cragg et al., Blood 101, 2003, pp. 1045-1052; and Cragg, Blood 103, 2004, pp. 2738-2743). Determination of FcRn binding and in vivo clearance/half-life can also be performed using methods known in the art (see, e.g., Petkova et al.,

. Immunol. 18(12), 2006, pp. 1759-1769).

Antibodies with reduced effector function include antibodies with the replacement of one or more of the following residues of the Fc region, such as 238, 265, 269, 270, 297, 327 and 329 (see, for example, US No. 6737056). These Fc mutants include Fc mutants with substitutions at two or more of the following amino acid positions: 265, 269, 270, 297, and 327, including the so-called "DANA" Fc region mutant carrying the substitution of residues 265 and 297 with alanine (US No. 7332581).

Some variants of antibodies with increased or decreased ability to bind to FcR are described (see, for example, US No. 6737056; WO 2004/056312 and Shields and others, J. Biol. Chem. 276, 2001, pp. 6591-6604).

In specific embodiments, the antibody variant comprises an Fc region with one or more amino acid substitutions that increase ADCC, such as substitutions at positions 298, 333, and/or 334 of the Fc region (EU residue numbering).

In some embodiments of the invention, changes are made in the Fc region that result in an altered (i.e., either increased or decreased) ability to bind C1q and/or complement dependent cytotoxicity (CDC), as described, for example, in US No. 6194551, WO 1999/51642 and Idusogie et al., J. Immunol. 164, 2000, pp. 4178-4184.

Antibodies with extended half-life and increased ability to bind to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgG to the embryo (Guyer et al., J. Immunol. 117, 1976, pp. 587-593 and Kim et al. , J. Immunol. 24, 1994, pp. 2429-2434), described in US 2005/0014934 A1 (Hinton et al.). These antibodies contain an Fc region with one or more substitutions that increase the binding of the Fc region to FcRn. These Fc region variants include variants with substitutions of one or more of the following Fc region residues: 378, 380, 382, 413, 424 or 434, for example, the replacement of residue 434 in the Fc region (US No. 7371826). Additional examples regarding other variants of the Fc region are also described in Duncan, Nature 322, 1988, ss. 738-740; in US Nos. 5648260 and 5624821 and in WO 1994/29351.

d) Cysteine-engineered antibody variants

In some embodiments, it may be desirable to create cysteine-engineered antibodies, such as "thioMAbs", in which one or more residues in the antibody are replaced with cysteine residues. In specific embodiments of the invention, the substituted residues are located at accessible sites on the antibody. By replacing these residues with cysteine, reactive thiol groups are placed at accessible sites on the antibody and can be used to conjugate the antibody to other moieties, such as drug moieties or drug linker moieties, to generate an immunoconjugate as indicated below in this description. In some embodiments, any one or more of the following residues may be replaced by a cysteine: V205 (Cabot numbering) light chain; A118 (EU number) of the heavy chain; and S400 (EU number) of the heavy chain Fc region. Antibodies constructed with cysteine can be generated according to the method, for example, described in US No. 7521541.

e) Antibody derivatives

In some embodiments, an antibody provided herein can be further modified to contain additional non-protein fragments known in the art and readily available. Fragments suitable for antibody derivatization include, but are not limited to, water-soluble polymers. Examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol/propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, copolymer ethylene/maleic anhydride, polyamino acids (either homopolymers or random copolymers) and dextran or poly(n-vinyl pyrrolidone) polyethylene glycol, propylene glycol homopolymers, propylene oxide/ethylene oxide copolymers, polyoxyethylated polyols (eg glycerol), polyvinyl alcohol and mixtures thereof. Polyethylene glycol propionaldehyde may have a manufacturing advantage due to its stability in water. The polymer may be of any molecular weight and may be branched or unbranched. The number of polymers attached to an antibody may vary, and if more than one polymer is attached, they may be the same or different molecules. In general, the amount and/or type of polymers used for derivatization can be determined with regard to, but not limited to, specific properties or functions of the antibody to be improved, whether the antibody derivative is intended to be used in therapy under certain conditions, etc.

Another embodiment of the invention are conjugates of an antibody and a non-protein fragment, which can be selectively heated by exposing it to radiation. In one embodiment, the non-protein moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102, 2005, pp. 11600-11605). The radiation can be of any wavelength and includes, but is not limited to, wavelengths that do not damage normal cells, but that heat the non-protein fragment to a temperature that kills cells closest to the conjugate: antibody-non-protein fragment.

8. Variants of Fc regions

One of the objects of the invention is an isolated polypeptide containing a variant of the Fc region with increased Fc-gamma RIIb-binding activity. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In some embodiments, a variant Fc region contains at least one amino acid residue change (e.g., a substitution) from the corresponding native Fc region sequence or a reference variant sequence (in some cases referred to collectively as the "parent" Fc region ). In some embodiments, a variant Fc region of the invention has increased binding activity for RIIb simian Fc-gamma compared to the parental Fc region. In specific embodiments, the RIIb simian Fc-gamma is Cynomolgus monkey Fc-gamma RIIb (SEQ ID NO: 223).

In some embodiments, the ratio [KD value of the parent Fc region for simian Fc-gamma RIIb)]/[KD value of the variant Fc region for simian Fc-gamma RIIb] may be 2.0 or more, 3.0 or more, 4.0 or more, 5.0 or more, 6.0 or more, 7.0 or more, 8.0 or more, 9.0 or more, 10 or more, 15 or more, 20 or more, 25 or more more, 30 or more, 40 or more, or 50 or more. In other embodiments, the Fc region variant has reduced binding activity to the simian Fc-gamma RIIIa. In some embodiments, the ratio [KD value of the parent Fc region for simian Fc-gamma RIIIa]/[KD value of the variant Fc region for simian Fc-gamma RIIIa] may be 0.50 or less, 0.40 or less, 0 .30 or less, 0.20 or less, 0.10 or less, 0.09 or less, 0.08 or less, 0.07 or less, 0.06 or less, 0.05 or less, 0.04 or less, 0.03 or less, 0.02 or less, or 0.01 or less. In some embodiments, the simian Fc-gamma RIIb has the sequence of SEQ ID NO: 223 (cynomolgus monkeys). In some embodiments, the simian Fc-gamma RIIIa has the sequence of SEQ ID NO: 224 (cynomolgus monkeys).

In other embodiments, the Fc region variant has increased binding activity to human Fc-gamma RIIb. In some embodiments, the ratio [KD value of parental Fc region for human Fc-gamma RIIb]/[KD value of variant Fc region for human Fc-gamma RIIb] may be 2.0 or more, 3.0 or more, 4 0 or more, 5.0 or more, 6.0 or more, 7.0 or more, 8.0 or more, 9.0 or more, 10 or more, 15 or more, 20 or more, 25 or more , 30 or more, 40 or more, or 50 or more. In other embodiments, the Fc region variant has reduced binding activity to human Fc-gamma RIIIa. In some embodiments, the ratio [KD value of parental Fc region for human Fc-gamma RIIIa]/[KD value of variant Fc region for human Fc-gamma RIIIa] may be 0.50 or less, 0.40 or less, 0 .30 or less, 0.20 or less, 0.10 or less, 0.09 or less, 0.08 or less, 0.07 or less, 0.06 or less, 0.05 or less, 0.04 or less, 0.03 or less, 0.02 or less, or 0.01 or less. In some embodiments, the human Fc-gamma RIIb has the sequence SEQ ID NO: 212, 213, or 214. In some embodiments, the human Fc-gamma RIIIa has the sequence SEQ ID NO: 215, 216, 217, or 218.

In other embodiments, the Fc region variant has reduced binding activity to human Fc-gamma RIIa (H-type) than to human Fc-gamma RIIb. In some embodiments, the ratio [KD value of parental Fc region for human Fc-gamma RIIa (H-type)]/[KD value of variant Fc region for human Fc-gamma RIIa (H-type)] may be 5.0 or less, 4.0 or less, 3.0 or less, 2.0 or less, 1.0 or less, 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less , 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, or 0.1 or less. In other embodiments, the Fc region variant has reduced binding activity to human Fc-gamma RIIa (R-type) than to human Fc-gamma RIIb. In some embodiments, the ratio [KD value of parental Fc region for human Fc-gamma RIIa R-type)]/[KD value of variant Fc region for human Fc-gamma RIIa (R-type)] may be 5.0 or less than, 4.0 or less, 3.0 or less, 2.0 or less, 1.0 or less, 0.9 or less, 0.8 or less, 0.7 or less, 0.6 or less, 0.5 or less, 0.4 or less, 0.3 or less, 0.2 or less, or 0.1 or less. In some embodiments, a human RIIa (H-type) Fc-gamma has the sequence of SEQ ID NO: 211. In some embodiments, a human RIIa (R-type) Fc-gamma has the sequence of SEQ ID NO: 210.

In some embodiments, the ratio [KD value of the parent Fc region for simian Fc-gamma RIIa]/[KD value of the variant Fc region for simian Fc-gamma RIIa] may be 2.0 or more, 3.0 or more, 4 0 or more, 5.0 or more, 6.0 or more, 7.0 or more, 8.0 or more, 9.0 or more, 10 or more, 15 or more, 20 or more, 25 or more , 30 or more, 40 or more, or 50 or more. In some embodiments, the simian Fc-gamma RIIa is selected from simian Fc-gamma RIIa1 (Fc-gamma RIIa1 cynomolgus (SEQ ID NO: 220)), simian Fc-gamma RIIa2 (e.g., Fc-gamma RIIa2 cynomolgus (SEQ ID NO: 221)) and simian Fc-gamma RIIa3 (eg Fc-gamma RIIa3 cynomolgus (SEQ ID NO: 222).

In another embodiment, the KD value of the Fc region variant for the simian Fc-gamma RIIb may be 1.0 x 10 ^-6 M or less, 9.0 x 10 ^-7 M or less, 8.0 x 10 ^-7 M, or less, 7.0×10 ^-7 M or less, 6.0×10 ^-7 M or less, 5.0×10 ^-7 M or less, 4.0×10 ^-7 M or less, 3.0×10 ^{- 7} M or less, 2.0×10 ^-7 M or less, or 1.0×10 ^-7 M or less. In another embodiment, the KD value of the Fc region variant for the simian Fc-gamma RIIIa may be 5.0 x 10 ^-7 M or more, 6.0 x 10 ^-7 M or more, 7.0 x 10 ^-7 M, or more, 8.0×10 ^-7 M or more, 9.0×10 ^-7 M or more, 1.0×10 ^-6 M or more, 2.0×10 ^-6 M or more, 3.0× 10 ^-6 M or more, 4.0×10 ^-6 M or more, 5.0×10 ^-6 M or more, 6.0×10 ^-6 M or more, 7.0×10 ^-6 M or more , 8.0×10 ^-6 M or more, 9.0×10 ^-6 M or more, or 1.0×10 ^-5 M or more. In another embodiment, the KD value of the Fc region variant for human Fc-gamma RIIb may be 2.0×10 ^-6 M or less, 1.0×10 ^-6 M or less, 9.0×10 ^-7 M, or less, 8.0×10 ^-7 M or less, 7.0×10 ^-7 M or less, 6.0×10 ^-7 M or less, 5.0×10 ^-7 M or less, 4.0× 10 ^-7 M or less, 3.0×10 ^-7 M or less, 2.0×10 ^-7 M or less, or 1.0×10 ^-7 M or less. In another embodiment, the KD value of the Fc region variant for human Fc-gamma RIIIa may be 1.0 x 10 ^-6 M or more, 2.0 x 10 ^-6 M or more, 3.0 x 10 ^-6 M, or more, 4.0×10 ^-6 M or more, 5.0×10 ^-6 M or more, 6.0×10 ^-6 M or more, 7.0×10 ^-6 M or more, 8.0× 10 ^-6 M or more, 9.0×10 ^-6 M or more, 1.0×10 ^-5 M or more, 2.0×10 ^-5 M or more, 3.0×10 ^-5 M or more , 4.0×10 ^-5 M or more or 5.0×10 ^-5 M or more. In another embodiment, the KD value of the Fc region variant for human Fc-gamma RIIa (H-type) may be 1.0×10 ^-7 M or more, 2.0×10 ^-7 M or more, 3.0× 10 ^-7 M or more, 4.0×10 ^-7 M or more, 5.0×10 ^-7 M or more, 6.0×10 ^-7 M or more, 7.0×10 ^-7 M or more , 8.0×10 ^-7 M or more, 9.0×10 ^-7 M or more, 1.0×10 ^-6 M or more, 2.0×10 ^-6 M or more, 3.0×10 ^-6 M or more, 4.0×10 ^-6 M or more, or 5.0×10 ^-6 M or more. In another embodiment, the KD value of the Fc region variant for a human Fc-gamma RIIa (R-type) may be 2.0×10 ^-7 M or more, 3.0×10 ^-7 M or more, 4.0× 10 ^-7 M or more, 5.0×10 ^-7 M or more, 6.0×10 ^-7 M or more, 7.0×10 ^-7 M or more, 8.0×10 ^-7 M or more , 9.0×10 ^-7 M or more, 1.0×10 ^-6 M or more, 2.0×10 ^-6 M or more, 3.0×10 ^-6 M or more, 4.0×10 ^-6 M or more or 5.0×10 ^-6 M or more.

In another embodiment, the KD value of the Fc region variant for the simian Fc-gamma RIIa may be 1.0×10 ^-6 M or less, 9.0×10 ^-7 M or less, 8.0×10 ^-7 M, or less, 7.0×10 ^-7 M or less, 6.0×10 ^-7 M or less, 5.0×10 ^-7 M or less, 4.0×10 ^-7 M or less, 3.0× 10 ^-7 M or less, 2.0×10 ^-7 M or less, or 1.0×10 ^-7 M or less. In some embodiments, the simian Fc-gamma RIIa can be selected from one of the following receptors: simian Fc-gamma RIIa1, simian Fc-gamma RIIa2, and simian Fc-gamma RIIa3.

When developing a pharmaceutical product for the treatment of diseases in humans, evaluating its efficacy and safety in monkeys is important because of their biological similarity to humans. With this in mind, the pharmaceutical product to be developed should preferably be cross-reactive to both humans and monkeys in terms of target binding activity.

"Fc-gamma receptors" (in the context of the present description, referred to as Fc-gamma receptors, Fc-gamma R or FcgR) are receptors that can bind to the Fc region of monoclonal antibodies of the isotype IgG1, IgG2, IgG3 and IgG4, and before in total is any member of the family of proteins encoded by the Fc-gamma receptor genes. In humans, this family includes, but is not limited to, Fc-gamma RI (CD64), including the isoforms Fc-gamma RIa, Fc-gamma Rib, and Fc-gamma RIc; Fc-gamma RII (CD32), including isoforms Fc-gamma RIIa (including allotypes H131 (type H) and R131 (type R)), Fc-gamma RIIb (including Fc-gamma RIIb-1 and Fc-gamma RIIb-2), and Fc-gamma RIIc; and Fc-gamma RIII (CD16), including the isoforms Fc-gamma RIIIa (including allotypes V158 and F158), and Fc-gamma RIIIb (including allotypes Fc-gamma RIIIb-NA1 and Fc-gamma RIIIb-NA2), and any human Fc-gamma R, Fc-gamma R isoforms or allotypes that have not yet been discovered. Fc-gamma RIIb1 and Fc-gamma RIIb2 are described as splicing variants of human Fc-gamma RIIb. In addition, a splicing variant has been described, designated Fc-gamma RIIb3 (J Exp Med, 170, 1989, pp. 1369-1385). In addition to these splicing variants, the human Fc-gamma RIIb includes all splicing variants registered with the NCBI, which are NP_001002273.1, NP_001002274.1, NP_001002275.1, NP_001177757.1 and NP_003992.3. In addition, human RIIb Fc-gamma includes each of the previously described gene polymorphisms as well as RIIb Fc-gamma (Arthritis Rheum. 48, 2003, pp. 3242-3252; Kono et al., Hum. Mol. Genet. 14, 2005 , pp. 2881-2892 and Kyogoju et al., Arthritis Rheum 46, 2002, pp. 1242-1254), and each gene polymorphism to be described in the future.

Fc-gamma RIIa has two allotypes, one in which the amino acid at position 131 of the Fc-gamma RIIa is histidine (type H) and the other in which the amino acid at position 131 is changed to arginine (type R) (Warrmerdam, J. Exp Med 172, 1990, pp 19-25).

Fc-gamma R includes but is not limited to Fc-gamma R derived from humans, mice, rats, rabbits and monkeys, and may be of any organism. Mouse Fc-gamma Rs include, but are not limited to, Fc-gamma RI (CD64), Fc-gamma RII (CD32), Fc-gamma RIII (CD16), and Fc-gamma RIII-2 (CD16-2) and any murine Fc-gamma R or Fc-gamma R isoforms. Unless specifically stated otherwise, the term "monkey Fc-gamma R" or a variation thereof refers to Fc-gamma RIIa1 (SEQ ID NO: 220), Fc-gamma RIIa2 ( SEQ ID NO: 221), Fc-gamma RIIa3 (SEQ ID NO: 222), Fc-gamma RIIb (SEQ ID NO: 223) or Fc-gamma RIIIaS (SEQ ID NO: 224) of cynomolgus monkeys.

The polynucleotide sequence of the human Fc-gamma RI is shown in SEQ ID NO: 199 (NM_000566.3); the polynucleotide sequence of human Fc-gamma RIIa is shown in SEQ ID NO: 200 (BC020823.1) or SEQ ID NO: 201 (NM_001136219.1); the polynucleotide sequence of human Fc-gamma RIIb is shown in SEQ ID NO: 202 (BC146678.1) or SEQ ID NO: 203 (NM_004001.3); the human Fc-gamma RIIIa polynucleotide sequence is shown in SEQ ID NO: 204 (BC033678.1) or SEQ ID NO: 205 (NM_001127593.1) and the human Fc-gamma RIIIb polynucleotide sequence is shown in SEQ ID NO: 206 (BC128562.1) .

The amino acid sequence of the human Fc-gamma RI is shown in SEQ ID NO: 207 (NP_000557.1); the amino acid sequence of human Fc-gamma RIIa is shown in SEQ ID NO: 208 (AAH20823.1), SEQ ID NO: 209, SEQ ID NO: 210, or SEQ ID NO: 211; the amino acid sequence of human Fc-gamma RIIb is shown in SEQ ID NO: 212 (AAI46679.1), SEQ ID NO: 213 or SEQ ID NO: 214; the amino acid sequence of human RIIIa Fc-gamma is shown in SEQ ID NO: 215 (AAH33678.1), SEQ ID NO: 216, SEQ ID NO: 217 or SEQ ID NO: 218 and the amino acid sequence of human RIIIb Fc-gamma is shown in SEQ ID NO : 219 (AAI28563.1).

The amino acid sequence of simian (cynomolgus) Fc-gamma RIIa is shown in SEQ ID NO: 220 (Fc-gamma RIIa1), SEQ ID NO: 221 (Fc-gamma RIIa2) or SEQ ID NO: 222 (Fc-gamma RIIa3); the amino acid sequence of simian (cynomolgus) Fc-gamma RIIb is shown in SEQ ID NO: 223 and the amino acid sequence of simian (cynomolgus) Fc-gamma RIIIa is shown in SEQ ID NO: 224.

One subject of the invention is polypeptides that contain variants of Fc regions with increased Fc-gamma RIIb-binding activity compared to the corresponding Fc-gamma RIIb-binding reference polypeptide. In other aspects of the invention, the polypeptide of the invention contains at least one amino acid change at least one position selected from the group consisting of: 231, 232, 233, 234, 235, 236, 237, 238, 239, 264 , 266, 267, 268, 271, 295, 298, 325, 326, 327, 328, 330, 331, 332, 334 and 396, according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

One of the objects of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity, which contain at least two amino acid changes, including: (a) one amino acid change at position 236 and (b) at least one amino acid change at at least one position selected from the group consisting of: 231, 232, 233, 234, 235, 237, 238, 239, 264, 266, 267, 268, 271, 295, 298, 325, 326, 327, 328, 330, 331, 332, 334, and 396, according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

One of the objects of the invention are polypeptides containing a variant of the Fc region with increased Fc-gamma RIIb-binding activity, which contains an amino acid change at position 236 according to the EU numbering.

One of the objects of the invention are polypeptides containing a variant of the Fc region with increased Fc-gamma RIIb-binding activity, which contains at least two amino acid changes, including: (a) one amino acid change at position 236 and (b) at least one amino acid change at at least one position selected from the group consisting of: 231, 232, 235, 239, 268, 295, 298, 326, 330 and 396, according to EU numbering. In another embodiment, the Fc region variant contains an amino acid change at at least one position selected from the group consisting of: 231, 232, 235, 239, 268, 295, 298, 326, 330, and 396, according to EU numbering . In a further embodiment of the invention, the variant Fc region contains an amino acid change at at least one position selected from the group consisting of: 268, 295, 326 and 330 according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus monkey sequence Cynomolgus monkey Fc-gamma RIIb (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

Another object of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity, which contain amino acid changes in one of the following positions (1)-(37): (1) positions 231, 236, 239, 268 and 330; (2) regulations 231, 236, 239, 268, 295 and 330; (3) provisions 231, 236, 268 and 330; (4) provisions 231, 236, 268, 295 and 330; (5) provisions 232, 236, 239, 268, 295 and 330; (6) provisions 232, 236, 268, 295 and 330; (7) provisions 232, 236, 268 and 330; (8) provisions 235, 236, 268, 295, 326 and 330; (9) provisions 235, 236, 268, 295 and 330; (10) provisions 235, 236, 268 and 330; (11) provisions 235, 236, 268, 330 and 396; (12) provisions 235, 236, 268 and 396; (13) provisions 236, 239, 268, 295, 298 and 330; (14) regulations 236, 239, 268, 295, 326 and 330; (15) provisions 236, 239, 268, 295 and 330; (16) provisions 236, 239, 268, 298 and 330; (17) provisions 236, 239, 268, 326 and 330; (18) provisions 236, 239, 268 and 330; (19) provisions 236, 239, 268, 330 and 396; (20) provisions 236, 239, 268 and 396; (21) provisions 236 and 268; (22) provisions 236, 268 and 295; (23) provisions 236, 268, 295, 298 and 330; (24) provisions 236, 268, 295, 326 and 330; (25) provisions 236, 268, 295, 326, 330 and 396; (26) provisions 236, 268, 295 and 330; (27) provisions 236, 268, 295, 330 and 396; (28) provisions 236, 268, 298 and 330; (29) provisions 236, 268, 298 and 396; (30) provisions 236, 268, 326 and 330; (31) regulations 236, 268, 326, 330 and 396; (32) provisions 236, 268 and 330; (33) provisions 236, 268, 330 and 396; (34) provisions 236, 268 and 396; (35) provisions 236 and 295; (36) provisions 236, 330 and 396; and (37) provisions 236 and 396, according to the EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In a further embodiment of the invention, the Fc region variant with enhanced Fc-gamma RIIb binding activity comprises at least one amino acid selected from the group consisting of: (a) Asp, Glu, Phe, Gly, His, Ile, Lys, Leu , Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, Tyr at position 231; (b) Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, Tyr at position 232; (c) Asp at position 233; (d) Trp, Tyr at position 234; (e) Trp at position 235; (f) Ala, Asp, Glu, His, Ile, Leu, Met, Asn, Gln, Ser, Thr, Val at position 236; (g) Asp, Tyr at position 237; (h) Glu, Ile, Met, Gln, Tyr at position 238; (i) Ile, Leu, Asn, Pro, Val at position 239; (j) Ile at position 264; (k) Phe at position 266; (m) Ala, His, Leu at position 267; (m) Asp, Glu at position 268; (o) Asp, Glu, Gly at position 271; (p) Leu at position 295; (p) Leu at position 298; (c) Glu, Phe, Ile, Leu at position 325; (r) Thr at position 326; (y) Ile, Asn at position 327; (f) Thr at position 328; (x) Lys, Arg at position 330; (c) Glu at position 331; (h) Asp at position 332; (w) Asp, Ile, Met, Val, Tyr at position 334; and (y) Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, Tyr at position 396; according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In a further embodiment of the invention, the Fc region variant with increased Fc-gamma RIIb binding activity comprises at least one amino acid change (eg, substitution) selected from the group consisting of: (a) Gly, Thr at position 231; (b) Asp at position 232; (c) Trp at position 235; (d) Asn, Thr at position 236; (e) Val at position 239; (e) Asp, Glu at position 268; (g) Leu at position 295; (h) Leu at position 298; (i) Thr at position 326; (j) Lys, Arg at position 330; and (k) Lys, Met at position 396 according to EU numbering. In a further embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb binding activity contains amino acid changes (e.g. substitutions): Asn at position 236, Glu at position 268, Lys at position 330 and Met at position 396 according to EU numbering . In a further embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb binding activity contains amino acid changes (eg substitutions): Asn at position 236, Asp at position 268 and Lys at position 330 according to EU numbering. In a further embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (e.g., substitutions): Asn at position 236, Asp at position 268, Leu at position 295, and Lys at position 330 according to EU numbering . In a further embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb binding activity contains amino acid changes (eg substitutions): Thr at position 236, Asp at position 268 and Lys at position 330 according to EU numbering. In a further embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb binding activity contains amino acid changes (e.g., substitutions): Asn at position 236, Asp at position 268, Leu at position 295, Thr at position 326, and Lys at position 330 according to EU-numbering. In a further embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb binding activity contains amino acid changes (e.g., substitutions): Trp at position 235, Asn at position 236, Asp at position 268, Leu at position 295, Thr at position 326, and Lys at position 330 according to the EU numbering.

One of the objects of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity, which contain an amino acid change at position 238 according to the EU numbering.

One of the objects of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity, which contain at least one amino acid change in at least one position selected from the group consisting of: 234, 238, 250, 264, 267, 307, and 330, according to the EU numbering. In other embodiments, the polypeptide contains at least one amino acid change at at least one position selected from the group consisting of: 234, 250, 264, 267, 307 and 330, according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

Another object of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity, which contain amino acid changes in one of the following positions (1)-(9): (1) positions 234, 238, 250, 307 and 330; (2) provisions 234, 238, 250, 264, 307 and 330; (3) provisions 234, 238, 250, 264, 267, 307 and 330; (4) provisions 234, 238, 250, 267, 307 and 330; (5) provisions 238, 250, 264, 307 and 330; (6) provisions 238, 250, 264, 267, 307 and 330; (7) provisions 238, 250, 267, 307 and 330; (8) provisions 238, 250 and 307; and (9) EU provisions 238, 250, 307 and 330. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In a further embodiment of the invention, the Fc region variant with increased Fc-gamma RIIb binding activity comprises at least one amino acid change (eg, substitution) selected from the group consisting of: (a) Tyr at position 234; (b) Asp at position 238; (c) Val at position 250, (d) Ile at position 264; (e) Ala at position 267; (f) Pro at position 307 and (g) Lys at position 330; according to EU numbering. In another embodiment of the invention, a variant of the Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (eg substitutions): Asp at position 238 according to EU numbering. In another embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (eg substitutions): Asp at position 238, Val at position 250, and Pro at position 307; according to EU numbering. In another embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (eg, substitutions): Asp at position 238, Val at position 250, Pro at position 307, and Lys at position 330; according to EU numbering. In another embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb binding activity contains amino acid changes (e.g., substitutions): Asp at position 238, Val at position 250, Ile at position 264, Pro at position 307, and Lys at position 330; according to EU numbering. In another embodiment, a variant Fc region with increased Fc-gamma RIIb binding activity contains amino acid changes (e.g., substitutions): Asp at position 238, Val at position 250, Ala at position 267, Pro at position 307, and Lys at position 307. position 330; according to EU numbering. In another embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (e.g., substitutions): Tyr at position 234, Asp at position 238, Val at position 250, Pro at position 307, and Lys at position 330; according to EU numbering. In another embodiment of the invention, a variant Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (e.g. substitutions): Tyr at position 234, Asp at position 238, Val at position 250, Ala at position 267, Pro at position 307 and Lys at position 330; according to EU numbering. In another embodiment of the invention, a variant of the Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (e.g. substitutions): Asp at position 238, Val at position 250, Ile at position 264, Ala at position 267, Pro at position 307 and Lys at position 330; according to EU numbering. In another embodiment of the invention, a variant of the Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (e.g. substitutions): Tyr at position 234, Asp at position 238, Val at position 250, Ile at position 264, Pro at position 307 and Lys at position 330; according to EU numbering. In another embodiment of the invention, a variant of the Fc region with increased Fc-gamma RIIb-binding activity contains amino acid changes (e.g. substitutions): Tyr at position 234, Asp at position 238, Val at position 250, Ile at position 264, Ala at position 267, Pro at position 307 and Lys at position 330; according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

Another object of the invention are selected polypeptides that contain variants of the Fc regions with increased isoelectric point (pI). In some embodiments, the Fc region variant described herein contains at least two amino acid changes in the parent Fc region. In some embodiments, each of the amino acid changes increases the isoelectric point (pI) of the variant Fc region relative to the parent Fc region. These polypeptides are based on the knowledge that elimination of an antigen from plasma can be accelerated by an antibody whose pI is increased by modifying at least two amino acid residues, for example when the antibody is administered in vivo.

According to the present invention pI can be either theoretical or experimentally determined pI. The value of pI can be determined, for example, using isoelectric focusing, known to experts in this field. The theoretical pI value can be calculated, for example, with software for gene and amino acid sequence analysis (Genetyx et al.).

In one of the embodiments of the invention, the pI value can be increased, for example, by at least 0.01, 0.03, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5 or more, at least 0.6, 0.7, 0.8, 0.9 or more, at least 1.0, 1.1, 1.2, 1.3, 1.4, 1, 5 or more, or at least 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 3.0, or more than the version before the modification.

или менее. Кроме того, методы определения областей, которые могут экспонироваться на поверхности, описаны у Pacios (Comput. Chem. 18(4), 1994, сс.377-386; J. Mol. Model. 1, 1995, cc. 46-53). На основе указанной выше информации можно отбирать пригодные аминокислотные остатки, локализованные на поверхности полипептида, который образует вариант Fc-области.In some embodiments, the pI-increasing amino acid may be displayed on the surface of a variant Fc region. According to the present invention, the amino acid that can be exposed on the surface, as a rule, is an amino acid residue localized on the surface of the polypeptide, forming a variant Fc region. An amino acid residue localized on the surface of a polypeptide means an amino acid residue whose side chain may be in contact with solvent molecules (most commonly water molecules). However, the side chain need not be wholly in contact with the solvent molecules, and even if only a portion of the side chain is in contact with the solvent molecules, the amino acid is considered to be a "surface-localized amino acid residue". Amino acid residues localized on the surface of a polypeptide also include amino acid residues localized near the surface, and which as a result can affect the electrical charge of another amino acid residue, even if the side chain of which is only partially in contact with solvent molecules. Those skilled in the art can create a homologous model of a polypeptide, for example, using commercially available software. Alternatively, methods known to those skilled in the art can be used, such as, for example, X-ray crystallography. The amino acid residues that can be exposed on the surface are determined, for example, using the coordinates of a three-dimensional model using a computer program such as the InsightII program (Accelrys). Sites that can be exposed on a surface can be determined using algorithms known in the art (for example, see Lee and Richards, J. Mol. Biol. 55, 1971, cc. 379-400; Connolly, J. Appl. Cryst 16, 1983, pp. 548-558). Sites that can be exposed on a surface can be determined with protein modeling software and 3D structure information. Suitable software for this purpose includes, for example, SYBYL Biopolymer Module software (Tripos Associates). When a user-defined "size" parameter is required in an algorithm, the "size" of the sample used in calculations can be specified as a radius of approximately

or less. In addition, methods for determining areas that can be exposed on a surface are described in Pacios (Comput. Chem. 18(4), 1994, pp. 377-386; J. Mol. Model. 1, 1995, pp. 46-53) . Based on the above information, suitable amino acid residues located on the surface of the polypeptide that form the variant Fc region can be selected.

In some embodiments, the polypeptide contains both a variant Fc region and an antigen-binding domain. In other embodiments, the antigen is a soluble antigen. In one embodiment, the antigen is present in the biological fluids (eg, plasma, interstitial fluid, lymphatic fluid, ascitic fluid, and pleural fluid) of individuals. The antigen may also be a membrane antigen.

In other embodiments of the invention, the antigen-binding activity of the antigen-binding domain varies depending on the level of ion concentration. In one embodiment, the ion concentration is not particularly limited, and means a hydrogen ion concentration (pH) or a metal ion concentration. In the context of the present description, metal ions are ions of group I elements excluding hydrogen, such as alkali metals and copper group elements, group II elements, such as alkaline earth metals and zinc group elements, group III elements excluding boron, group IV elements excluding carbon and silicon, group VIII elements such as iron group and platinum group elements, elements belonging to subgroup A of groups V, VI and VII, and metal elements such as antimony, bismuth and polonium. According to the present invention, metal ions include, for example, calcium ions as described in WO 2012/073992 and WO 2013/125667. In one of the embodiments of the invention, the "ion concentration level" can be a level that affects the biological behavior of the antigen-binding domain, located between a low concentration of ions and a high concentration of ions. In addition, the term "antigen-binding activity of antigen-binding domains varies depending on the level of ion concentration" means that the antigen-binding activity of antigen-binding domains changes depending on whether the ion concentration is low or high (wherein the antigen-binding domain is referred to as "ion concentration-dependent antigen-binding domain"). domain"). The antigen-binding activity of antigen-binding domains at a high level of ion concentration may be higher (stronger) or lower (weaker) than at a low level of ion concentration. In one embodiment, ion concentration dependent antigen binding domains (such as pH dependent antigen binding domains or calcium ion concentration dependent antigen binding domains) can be obtained using known methods, such as those described in WO 2009/125825, WO 2012/073992 and WO 2013/046722.

According to the present invention, the antigen-binding activity of an antigen-binding domain at a high level of calcium ion concentration may be higher than at a low level of calcium ion concentration. The high calcium ion concentration is not limited to specific values, but may be a concentration selected from the range of 100 µM to 10 mM, 200 µM to 5 mM, 400 µM to 3 mM, 200 µM to 2 mM, 400 µM to 1 mm or from 500 μm to 2.5 mm, which is preferred due to the proximity to the concentration of calcium ions in plasma (blood) in vivo. In addition, the low calcium ion concentration is not limited to specific values, but may be a concentration selected from the range of 0.1 to 30, 0.2 to 20, 0.5 to 10, 1 to 5, or 2 to 4 μM, which is preferred due to its proximity to the concentration of calcium ions in early endosomes in vivo.

In one embodiment, the ratio between antigen-binding activities at a low calcium ion level and a high calcium ion level is not limited to specific values, but the ratio of the dissociation constant (KD) at a low calcium ion level and KD at a high calcium ion level, t .e. KD (low calcium ion level)/KD (high calcium ion level) is 2 or more, 10 or more, or 40 or more. The upper limit of the ratio may be 400, 1000 or 10000 if said antigen-binding domain can be obtained using techniques known to those skilled in the art. Alternatively, for example, the dissociation reaction rate constant (kd) can be used instead of KD. In this case, the ratio of kd at a low level of calcium ion concentration and kd at a high level of calcium ion concentration, i.e. kd (low calcium ion level)/kd (high calcium ion level) is 2 or more, 5 or more, 10 or more, or 30 or more. The upper limit of the ratio may be 50, 100 or 200 if said antigen-binding domain can be obtained using techniques known to those skilled in the art.

According to the present invention, the antigen-binding activity of an antigen-binding domain at a low concentration of hydrogen ions (at neutral pH) can be higher than at a high concentration of hydrogen ions (at acidic pH). The acidic pH may be, for example, a pH selected from a pH range of 4.0 to 6.5, 4.5 to 6.5, 5.0 to 6.5, or 5.5 to 6.5, which is preferred due to proximity to in vivo pH in early endosomes. The acidic pH may also be, for example, pH 5.8 or pH 6.0. In specific embodiments, the acidic pH is pH 5.8. In addition, the neutral pH may be, for example, a pH selected from a pH range of 6.7 to 10.0, 6.7 to 9.5, 7.0 to 9.0, or 7.0 to 8 0, which is preferred due to its proximity to the in vivo pH in plasma (blood). Neutral pH can also be, for example, pH 7.4 or pH 7.0. In a specific embodiment of the invention, the neutral pH is pH 7.4.

In one embodiment of the invention, the ratio between antigen-binding activities under acidic pH conditions and under neutral pH conditions is not limited to specific values, but the ratio of dissociation constant (KD) under acidic pH conditions and KD under neutral pH conditions, i.e. KD (acidic pH conditions)/KD (neutral pH conditions) is 2 or more, 10 or more, or 40 or more. The upper limit of the ratio may be 400, 1000 or 10000 if said antigen-binding domain can be obtained using techniques known to those skilled in the art. Alternatively, for example, the dissociation reaction rate constant (kd) can be used instead of KD. In this case, the ratio of kd under acidic pH conditions and kd under neutral pH conditions, i.e. kd (acidic pH conditions)/kd (neutral pH conditions) is 2 or more, 5 or more, 10 or more, or 30 or more. The upper limit of the ratio may be 50, 100 or 200 if said antigen-binding domain can be obtained using techniques known to those skilled in the art.

In one embodiment, for example, at least one amino acid residue is replaced with an amino acid residue with a side chain pKa of 4.0-8.0 and/or at least one amino acid residue with a side chain pKa of 4.0-8.0 is inserted into an antigen binding domain as described in WO 2009/125825. An amino acid may be substituted and/or inserted at any site if the antigen-binding activity of the antigen-binding domain becomes weaker under acidic pH conditions compared to neutral pH conditions relative to the variant prior to the substitution or insertion. When the antigen binding domain has a variable region or CDR, the site may be within the variable region or CDR. Those skilled in the art can appropriately determine the amount of amino acids substituted or inserted; and this number can be 1 or more. Amino acids with a side chain pKa of 4.0-8.0 can be used to alter the antigen-binding activity of the antigen-binding domain depending on the level of hydrogen ion concentration. These amino acids include, for example, naturally occurring amino acids such as His (H) and Glu (E), and non-naturally occurring amino acids such as histidine analogs (US 2009/0035836), m-NO ₂ -Tyr ( pKa 7.45), 3.5-Br ₂ -Tyr (pKa 7.21), and 3.5-I ₂ -Tyr (pKa 7.38) (Heyl et al., Bioorg. Med. Chem. 11( 17), 2003, pp. 3761-3768). It is also possible to use amino acids with a side chain pKa of 6.0-7.0, which include, for example, His (H).

In another embodiment, the preferred antigen binding domains for the elevated pI variant Fc region are known and can be obtained using the methods described in Japanese Patent Applications JP 2015-021371 and JP 2015-185254.

In some embodiments, the elevated pI variant Fc region contains at least two amino acid changes at at least two positions selected from the group consisting of: 285, 311, 312, 315, 318, 333, 335, 337, 341 , 342, 343, 384, 385, 388, 390, 399, 400, 401, 402, 413, 420, 422, and 431, according to EU numbering.

In other embodiments, the elevated pI variant Fc region contains at least two amino acid changes at at least two positions selected from the group consisting of: 311, 341, 343, 384, 399, 400, 401, 402, and 413, according to EU numbering.

Another object of the invention are polypeptides containing variants of Fc regions with increased pI, which contain amino acid changes in one of the following positions (1)-(10): (1) positions 311 and 341; (2) provisions 311 and 343; (3) provisions 311, 343 and 413; (4) provisions 311, 384 and 413; (5) provisions 311 and 399; (6) provisions 311 and 401; (7) provisions 311 and 413; (8) provisions 400 and 413; (9) provisions 401 and 413; and (10) provisions 402 and 413; according to EU numbering.

The method for reducing the pI of a protein involves, for example, reducing the amount of amino acids with a negatively charged side chain (for example, aspartic acid and glutamic acid) and/or increasing the amount of amino acids with a positively charged side chain (for example, arginine, lysine and histidine) under neutral pH conditions. Amino acids with a negatively charged side chain have a negative charge corresponding to -1 under pH conditions, which is well above the pKa value of the side chain, according to theory well known to those skilled in the art. For example, the theoretical pKa for the side chain of aspartic acid is 3.9, and the side chain has a negative charge corresponding to -1 under neutral pH conditions (eg, in a solution of pH 7.0). Conversely, amino acids with a positively charged side chain have a positive charge corresponding to +1 under pH conditions that are sufficiently lower than the pKa value of their side chain. For example, the theoretical pKa value for the arginine side chain is 12.5, and the side chain has a positive charge corresponding to +1 under neutral pH conditions (eg, pH 7.0 solution). Moreover, as is known, amino acids, the side chains of which do not have a charge under neutral pH conditions (for example, in a solution with pH 7.0), include 15 types of naturally occurring amino acids, i.e. alanine, cysteine, phenylalanine, glycine, isoleucine, leucine, methionine, asparagine, proline, glutamine, serine, threonine, valine, tryptophan and tyrosine. It goes without saying that the amino acids that can be used to increase pI may be non-naturally occurring amino acids.

Based on the above, using the method of increasing the pI of a protein under neutral pH conditions (for example, in a solution with pH 7.0), it is possible to change the charge of the protein of interest by +1, for example, by replacing amino acids with uncharged side chains of aspartic acid or glutamic acid (whose side chain has a negative charge of -1) in the amino acid sequence of a protein. In addition, a +1 charge change can be effected in a protein, for example, by substituting arginine or lysine (whose side chain has a +1 positive charge) for amino acids with uncharged side chains. In addition, a +2 charge change of the protein can be carried out simultaneously by substituting arginine or lysine (whose side chain has a +1 positive charge) for aspartic acid or glutamic acid (whose side chain has a negative charge of -1). Alternatively, to increase the pI of a protein, amino acids with uncharged side chains and/or preferably amino acids with positively charged side chains can be added or inserted into the amino acid sequence of the protein, or amino acids with uncharged side chains and/or preferably amino acids with negatively charged side chains present in the amino acid sequence of a protein can be removed by deletion. Obviously, for example, the N-terminal and C-terminal amino acid residues of a protein have a charge inherent in their main chain (NH ₃ ⁺ amino groups at the N-terminus and COO ^- carbonyl group at the C-terminus) in addition to the charges inherent in their side chain. Thus, the pI of a protein can be increased by certain addition, deletion, substitution or insertion in the functional groups of the main chain.

Substituting an amino acid to increase pI includes, for example, substituting an amino acid with an uncharged side chain for an amino acid with a negatively charged side chain, substituting an amino acid with a positively charged side chain for an amino acid with an uncharged side chain, and substituting an amino acid with a positively charged side chain for an amino acid with a negatively charged side chain in an amino acid. the sequence of the parent Fc region, which is carried out individually or in appropriate combination.

Insertion or addition of an amino acid to increase pI includes, for example, insertion or addition of an amino acid with an uncharged side chain and/or insertion or addition of an amino acid with a positively charged side chain into the amino acid sequence of the parent Fc region, which is carried out individually or in an appropriate combination.

Deletion of an amino acid to increase pI includes, for example, deletion of an amino acid with an uncharged side chain and/or deletion of an amino acid with a negatively charged side chain in the amino acid sequence of the parent Fc region, which is carried out individually or in an appropriate combination.

In one embodiment, the naturally occurring amino acids that are used to increase pI can be classified as follows: (a) the negatively charged side chain amino acid can be Glu (E) or Asp (D); (b) the uncharged side chain amino acid may be Ala (A), Asn (N), Cys (C), Gln (Q), Gly (G), His (H), Ile (I), Leu (L) , Met (M), Phe (F), Pro (P), Ser (S), Thr (T), Trp (W), Tyr (Y), or Val (V); and (c) the positively charged side chain amino acid may be His (H), Lys (K), or Arg (R). In one embodiment of the invention, after modification by insertion or substitution, the amino acid is Lys (K) or Arg (R).

Another object of the invention are isolated polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity and increased pI. In some embodiments, the Fc region variant described herein contains at least two amino acid changes in the parent Fc region.

One of the objects of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity and increased pI, which contain at least three amino acid changes, which are: (a) at least one amino acid change in at least one position selected from the group consisting of: 231, 232, 233, 234, 235, 236, 237, 238, 239, 264, 266, 267, 268, 271, 295, 298, 325, 326, 327, 328, 330, 331, 332, 334 and 396, according to EU numbering, and (b) at least two amino acid changes at least two positions selected from the group consisting of: 285, 311, 312, 315, 318, 333 , 335, 337, 341, 342, 343, 384, 385, 388, 390, 399, 400, 401, 402, 413, 420, 422 and 431, according to EU numbering.

One of the objects of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity and increased pI, which contain at least three amino acid changes, which are: (a) at least one amino acid change in at least one position selected from the group consisting of: 231, 232, 235, 236, 239, 268, 295, 298, 326, 330, and 396, according to EU numbering, and (b) at least two amino acid changes in at least least in two positions selected from the group consisting of: 311, 341, 343, 384, 399, 400, 401, 402 and 413, according to EU numbering.

Another object of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity and increased pI, which contain amino acid changes in one of the following positions (1)-(9): (1) positions 235, 236, 268 , 295, 311, 326, 330 and 343; (2) regulations 236, 268, 295, 311, 326, 330 and 343; (3) provisions 236, 268, 295, 311, 330 and 413; (4) regulations 236, 268, 311, 330, 396 and 399; (5) provisions 236, 268, 311, 330 and 343; (6) provisions 236, 268, 311, 330, 343 and 413; (7) provisions 236, 268, 311, 330, 384 and 413; (8) provisions 236, 268, 311, 330 and 413; and (9) regulations 236, 268, 330, 396, 400 and 413; according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

One of the objects of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity and increased pI, which contain at least three amino acid changes, which are: (a) at least one amino acid change in at least one position selected from the group consisting of: 234, 238, 250, 264, 267, 307 and 330, and (b) at least two amino acid changes at least two positions selected from the group consisting of: 285, 311, 312, 315, 318, 333, 335, 337, 341, 342, 343, 384, 385, 388, 390, 399, 400, 401, 402, 413, 420, 422, and 431, according to EU numbering. In other embodiments, the polypeptides contain at least two amino acid changes at least two positions selected from the group consisting of: 311, 341, 343, 384, 399, 400, 401, 402, and 413, according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

Another object of the invention are polypeptides containing variants of Fc regions with increased Fc-gamma RIIb-binding activity and increased pI, which contain amino acid changes in one of the following positions (1)-(16): (1) positions 234, 238, 250 , 264, 307, 311, 330 and 343; (2) regulations 234, 238, 250, 264, 307, 311, 330 and 413; (3) provisions 234, 238, 250, 264, 267, 307, 311, 330 and 343; (4) provisions 234, 238, 250, 264, 267, 307, 311, 330 and 413; (5) provisions 234, 238, 250, 267, 307, 311, 330 and 343; (6) regulations 234, 238, 250, 267, 307, 311, 330 and 413; (7) provisions 234, 238, 250, 307, 311, 330 and 343; (8) provisions 234, 238, 250, 307, 311, 330 and 413; (9) provisions 238, 250, 264, 267, 307, 311, 330 and 343; (10) provisions 238, 250, 264, 267, 307, 311, 330 and 413; (11) regulations 238, 250, 264, 307, 311, 330 and 343; (12) regulations 238, 250, 264, 307, 311, 330 and 413; (13) regulations 238, 250, 267, 307, 311, 330 and 343; (14) provisions 238, 250, 267, 307, 311, 330 and 413; (15) provisions 238, 250, 307, 311, 330 and 343; and (16) EU provisions 238, 250, 307, 311, 330 and 413.

In a further embodiment of the invention, the Fc region variant comprises amino acid changes selected from any of the single changes, combinations of single changes, or combinations of changes shown in Tables 14-30.

In some embodiments, the polypeptide comprises a variant Fc region of the present invention. In a further embodiment, the polypeptide is an antibody heavy chain constant region. In a further embodiment of the invention, the polypeptide is an antibody heavy chain. In a further embodiment of the invention, the polypeptide is an antibody. In a further embodiment, the polypeptide is an Fc-containing fusion protein.

Another embodiment of the invention is a polypeptide containing one of the amino acid sequences of SEQ ID NO: 229-381.

In the context of the present description, the term "parent Fc-region" refers to the Fc-region before the introduction of the amino acid(s) change(s) specified(s) in the present description. Preferred examples of a parental Fc region are Fc regions from native antibodies. Antibodies include, for example, IgA (IgA1, IgA2), IgD, IgE, IgG (IgG1, IgG2, IgG3, IgG4) and IgM or the like. The antibodies may be of human or simian origin (eg cynomolgus, rhesus monkey, marmoset, chimpanzee or baboon). Native antibodies may also include naturally occurring mutations. A variety of IgG allotypic sequences associated with genetic polymorphisms are described in Sequences proteins of immunological interest), NIH Publication No. 91-3242, and any of them can be used in the present invention. In particular, in the case of human IgG1, the amino acid sequence at positions 356-358 (EU numbering) can be either DEL or EEM. Preferred examples of the parental Fc region include Fc regions from the heavy chain constant region of human IgG1 (SEQ ID NO: 195), human IgG2 (SEQ ID NO: 196), human IgG3 (SEQ ID NO: 197), and human IgG4 (SEQ ID NO: 198). Another preferred example of a parental Fc region is the Fc region from the SG1 heavy chain constant region (SEQ ID NO: 9). In addition, the parent Fc region may be an Fc region obtained by adding an amino acid(s) change(s) other than the amino acid(s) change(s) specified(s) in the present description for Fc- regions from the native antibody.

и др., J. Biol. Chem. 281(33), 2006, сс. 23514-23524; Petkova и др., Intl. Immunol. 18(12), 2006, сс. 1759-1769; Zalevsky и др., Nat. Biotechnol. 28(2), 2010, сс. 157-159; WO 2006/019447; WO 2006/053301; и WO 2009/086320), и аминокислотные замены, которые повышают гетерогенность или стабильность антитела (WO 2009/041613). Альтернативно этому, полипептиды, обладающие способностью усиливать клиренс антигена, которые описаны в WO 2011/122011, WO 2012/132067, WO 2013/046704 или WO 2013/180201, полипептиды, обладающие способностью специфически связываться с тканью-мишенью, которые описаны в WO 2013/180200, полипептиды, обладающие способностью повторно связываться с множеством молекул антигенов, которые описаны в WO 2009/125825, WO 2012/073992 или WO 2013/047752, можно объединять с вариантом Fc-области, указанном в настоящем описании. Альтернативно этому, с целью придания способности связываться с другими антигенами аминокислотные изменения, описанные в ЕР 1752471 и ЕР 1772465, можно объединять в СН3 варианта Fc-области, указанного в настоящем описании. Альтернативно этому, с целью повышения удерживания в плазме можно объединять аминокислотные изменения, которые снижают pI константной области (WO 2012/016227), в варианте Fc-области, указанном в настоящем описании. Альтернативно этому, с целью повышения поглощения клеткой можно объединять аминокислотные изменения, которые повышают pI константной области (WO 2014/145159), в варианте Fc-области, указанном в настоящем описании. Альтернативно этому, с целью усиления элиминации молекулы-мишени из плазмы можно объединять аминокислотные изменения, которые повышают pI константной области (заявки на японский патент JP 2015-021371 и JP 2015-185254), в варианте Fc-области, указанном в настоящем описании. В одном из вариантов осуществления изобретения указанное изменение может включать, например, замену по меньшей мере одного положения, выбранного из группы, состоящей из 311, 343, 384, 399, 400 и 413 согласно EU-нумерации. В другом варианте осуществления изобретения указанная замена может представлять собой замену аминокислоты на Lys или Arg в каждом положении.In addition, amino acid changes that are made for other purpose(s) can be combined in the Fc region variant described herein. For example, amino acid substitutions can be added that increase FcRn binding activity (Hinton et al., J. Immunol. 176(1), 2006, pp. 346-356;

et al., J. Biol. Chem. 281(33), 2006, pp. 23514-23524; Petkova et al., Intl. Immunol. 18(12), 2006, pp. 1759-1769; Zalevsky et al., Nat. Biotechnol. 28(2), 2010, pp. 157-159; WO 2006/019447; WO2006/053301; and WO 2009/086320), and amino acid substitutions that increase antibody heterogeneity or stability (WO 2009/041613). Alternatively, polypeptides having the ability to enhance antigen clearance as described in WO 2011/122011, WO 2012/132067, WO 2013/046704 or WO 2013/180201, polypeptides having the ability to specifically bind to a target tissue as described in WO 2013 /180200, polypeptides having the ability to re-bind to multiple antigen molecules as described in WO 2009/125825, WO 2012/073992 or WO 2013/047752 can be combined with the Fc region variant described herein. Alternatively, in order to confer the ability to bind to other antigens, the amino acid changes described in EP 1752471 and EP 1772465 can be combined into the CH3 variant of the Fc region specified in the present description. Alternatively, amino acid changes that reduce the pI of the constant region (WO 2012/016227) in the Fc region variant described herein can be combined to increase plasma retention. Alternatively, amino acid changes that increase the pI of the constant region (WO 2014/145159), in the Fc region variant described herein, can be combined to increase cellular uptake. Alternatively, in order to enhance elimination of the target molecule from plasma, amino acid changes that increase the pI of the constant region (Japanese Patent Applications JP 2015-021371 and JP 2015-185254), in the Fc region variant described herein, can be combined. In one of the embodiments of the invention, the specified change may include, for example, the replacement of at least one position selected from the group consisting of 311, 343, 384, 399, 400 and 413 according to the EU numbering. In another embodiment of the invention, said substitution may be an amino acid substitution with Lys or Arg at each position.

и др., J. Biol. Chem. 281, 2006, cc. 23514-23524); замену на Gln Thr в положении 250 и замену на Leu Met в положении 428 (Hinton и др., J. Immunol. 176 (1), 2006, сс. 346-356); замену на His Asn в положении 434 (Zheng и др., Clin. Pharmacol. Ther. 89 (2), 2011, сс. 283-290), и изменения, описанные в WO 2010/106180, WO 2010/045193, WO 2009/058492, WO 2008/022152, WO 2006/050166, WO 2006/053301, WO 2006/031370, WO 2005/123780, WO 2005/047327, WO 2005/037867, WO 2004/035752 или WO 2002/060919.Такие изменения могут включать, например, по меньшей мере одно изменение, выбранное из группы, состоящей из замены на Leu Met в положении 428, замены на Ala Asn в положении 434 и замены на Thr Tyr в положении 436. Указанные изменения включают также замену на Arg Gln в положении 438 и/или замену на Glu Ser в положении 440 (заявки на японский патент JP 2015-021371 и JP 2015-185254).Amino acid changes resulting in increased ability to bind to human FcRn at acidic pH can also be combined in the Fc region variant described herein. In particular, such changes may include, for example, a change to Leu Met at position 428 and a change to Ser Asn at position 434, according to EU numbering (Zalevsky et al., Nat. Biotechnol. 28, 2010, pp. 157-159) ; substitution for Ala Asn at position 434 (Deng et al., Metab. Dispos. 38(4), 2010, pp. 600-605); a Tyr Met substitution at position 252, a Thr Ser substitution at position 254, and a Glu Thr substitution at position 256 (

et al., J. Biol. Chem. 281, 2006, pp. 23514-23524); a change to Gln Thr at position 250 and a change to Leu Met at position 428 (Hinton et al., J. Immunol. 176 (1), 2006, pp. 346-356); substitution for His Asn at position 434 (Zheng et al., Clin. Pharmacol. Ther. 89 (2), 2011, pp. 283-290), and changes described in WO 2010/106180, WO 2010/045193, WO 2009 /058492, WO 2008/022152, WO 2006/050166, WO 2006/053301, WO 2006/031370, WO 2005/123780, WO 2005/047327, WO 2005/037867, WO 2004/03.WO5602 may include, for example, at least one change selected from the group consisting of a change to Leu Met at position 428, a change to Ala Asn at position 434, and a change to Thr Tyr at position 436. These changes also include a change to Arg Gln at position 436. position 438 and/or a change to Glu Ser at position 440 (Japanese patent applications JP 2015-021371 and JP 2015-185254).

Two or more polypeptides containing an Fc region variant as described herein may be included in a single molecule in which the two Fc region variant polypeptides are in an association that closely resembles that of an antibody. The type of antibody is not limited, and IgA (IgA1, IgA2), IgD, IgE, IgG (IgG1, IgG2, IgG3, IgG4) and IgM or the like can be used.

Two associated polypeptides containing variants of the Fc regions may be polypeptides that contain variants of the Fc regions into which the same amino acid change(s) has been introduced (hereinafter referred to as homologous variants in the context of the present description). Fc regions), or polypeptides that contain variants of Fc regions into which different amino acid(s) change(s) have been introduced, or alternatively, polypeptides that contain variants of Fc regions, characterized in that the amino acid ( s) the change(s) is(are) introduced into only one of the Fc regions ((hereinafter referred to as heterologous polypeptides of Fc region variants). One of the preferred amino acid changes is a change in the loop structure from position 233 to 239 ( EU numbering) in the CH2 domain of the Fc region which is involved in binding to Fc-gamma RIIb and Fc-gamma RIIa Preferably a change that increases Fc-gamma RIIb-c binding activity and/or selectivity is introduced into the loop structure of the CH2 domain of one of the Fc regions, and another change is introduced into the loop structure of the CH2 domain of the other Fc region, which destabilizes it. Examples of amino acid changes that can destabilize the loop structure of the CH2 domain may be the substitution of at least one amino acid selected from the amino acids at positions 235, 236, 237, 238 and 239 with a different amino acid. In particular, destabilization can be carried out, for example, by changing the amino acid at position 235 to Asp, Gln, Glu or Thr, changing the amino acid at position 236 to Asn, changing the amino acid at position 237 to Phe or Trp, changing the amino acid at position 238 to Glu, Gly or Asn and amino acid changes at position 239 to Asp or Glu, according to EU numbering.

For the association of heterologous polypeptides containing variant Fc regions, the technique of suppressing unwanted association of homologous polypeptides containing variant Fc regions by introducing electrostatic repulsion into the interface of the CH2 or CH3 domain of the Fc region described in WO 2006/106905 can be used.

Examples of amino acid residues contacting at the interface of the CH2 or CH3 domain of the Fc region include residue at position 356 (EU numbering), residue at position 439 (EU numbering), residue at position 357 (EU numbering), residue at position 370 (EU numbering), a remainder at position 399 (EU numbering), and a remainder at position 409 (EU numbering) in the CH3 domain.

More specifically, for example, it is possible to obtain an Fc region in which 1-3 pairs of amino acid residues selected from (1) to (3) below have the same charge: (1) amino acid residues at positions 356 and 439 (EU -numbering) in the CH3 domain; (2) amino acid residues at positions 357 and 370 (EU numbering) in the CH3 domain; and (3) amino acid residues at positions 399 and 409 (EU numbering) in the CH3 domain.

In addition, heterologous polypeptides can be prepared containing Fc region variants in which 1-3 pairs of amino acid residues selected from (1) to (3) above have the same charge in the CH3 domain of the first Fc region, and pairs of amino acid residues selected in the above first Fc region also have the same charge in the CH3 domain of the second Fc region, provided that the charges in the first and second Fc regions are opposite.

In the above Fc regions, for example, negatively charged amino acid residues are preferably selected from glutamic acid (E) and aspartic acid (D), and positively charged amino acid residues are preferably selected from lysine (K), arginine (R) and histidine (H).

Other known techniques can also be used to associate heterologous polypeptides containing Fc region variants. In particular, this technique is carried out by replacing the side chain of an amino acid in one of the Fc regions with a larger side chain (protrusion; which is referred to as "bulge") and by replacing the side chain of an amino acid present in the Fc region with a smaller side chain ( cavity; which is referred to as the "cavity"), placing the protrusion in the cavity. This can facilitate efficient association between Fc region-containing polypeptides that have different amino acid sequences from each other (WO 1996/027011; Ridgway et al., Prot. Eng. 9, 1996, pp. 617-621; Merchant et al., Nat Biotech 16, 1998, pp. 677-681).

In addition, other known techniques can also be used to associate heterologous polypeptides containing variant Fc regions. The association of polypeptides containing an Fc region can be efficiently induced using a chain-exchange generated CH3 domain of heterodimers (Davis et al., Prot. Eng. Des. & Sel., 23, 2010, pp. 195-202). This technique can also be used to efficiently induce association between Fc region-containing polypeptides with different amino acid sequences.

In addition, methods for producing heterodimerized antibodies that use the association of CH1 and CL antibodies and the association of VH and VL described in WO 2011/028952 can also be used.

Similar to the method described in WO 2008/119353 and WO 2011/131746, it is also possible to apply the technique of obtaining heterodimerized antibodies by first creating two types of homodimerized antibodies, incubating the antibodies under reducing conditions to dissociate them and allowing them to re-associate.

Similar to the method described by Strop, J. Mol. Biol. 420, 2012, pp. 204-219, a technique for making heterodimerized antibodies by introducing charged residues such as Lys, Arg, Glu, and Asp can also be used to introduce electrostatic repulsion into the CH3 domains.

In addition, similar to the method described in WO 2012/058768, you can also apply the method of obtaining heterodimerized antibodies by introducing changes in the CH2 and CH3 domains.

When two polypeptides containing an Fc region variant that have different amino acid sequences are simultaneously expressed to produce polypeptides containing heterologous Fc region variants, polypeptides containing homologous Fc region variants are generally formed as impurities. In such cases, polypeptides containing heterologous Fc region variants can be efficiently obtained by separating them and purifying them from polypeptides containing homologous Fc region variants using known techniques. A method for efficiently separating and purifying heterodimerized antibodies from homodimerized antibodies based on the use of ion exchange chromatography by introducing amino acid changes into the variable regions of two types of antibody heavy chains to create a difference in isoelectric points between homodimerized antibodies and heterodimerized antibodies is described (WO 2007/114325). Another method is described for purifying heterodimerized antibodies using protein A chromatography by constructing a heterodimerized antibody containing two types of heavy chains from mouse IgG2a that bind to protein A and rat IgG2b that does not bind to protein A (WO 1998/050431 and WO 1995/ 033844).

In addition, a heterodimerized antibody can be efficiently purified by protein A chromatography by replacing the amino acid residues at positions 435 and 436 (EU numbering), which are located at the protein A binding site in the heavy chain of the antibody, with amino acids such as Tyr or His , resulting in different binding affinities for protein A.

In the present invention, an amino acid change means any of such changes as substitution, deletion, addition, insertion and modification, or a combination thereof. In the present invention, an amino acid change may be referred to as an amino acid mutation.

When substituting amino acid residues, substitution with a different amino acid residue can be carried out to change the characteristics indicated in subparagraphs (a) to (c) below: (a) the structure of the polypeptide backbone of the region of the folded structure or helical structure; (b) electrical charge or hydrophobicity at the target site; or (c) side chain size.

Amino acid residues are classified into the following groups based on common side chain properties: (a) hydrophobic: norleucine, Met, Ala, Val, Leu and Ile; (b) neutral hydrophilic: Cys, Ser, Thr, Asn and Gln; (c) acidic: Asp and Glu; (d) main: His, Lys and Arg; (e) residues that affect chain orientation: Gly and Pro; and (e) aromatic: Trp, Tyr and Phe.

Amino acid changes are obtained using various methods known to those skilled in the art. These methods include, but are not limited to, the site-directed mutagenesis method (Hashimoto-Gotoh et al., Gene 152, 1995, pp. 271-275; Zoller, Meth. Enzymol. 100, 1983, pp. 4 68-500; Kramer et al., Nucleic Acids Res. 12, 1984, pp. 9441-9456 Kramer and Fritz, Methods Enzymol 154, 1987, pp. 350-367 and Kunkel, Proc. Natl. Acad. Sci. USA 82, 1985 , pp. 488-492), a method based on PCR mutagenesis and cassette mutagenesis.

The number of amino acid changes introduced from the Fc region is not limited. In some embodiments, it may be 1, 2 or less, 3 or less, 4 or less, 5 or less, 6 or less, 8 or less, 10 or less, 12 or less, 14 or less, 16 or less, 18 or less or 20 or less.

Amino acid modification includes post-translational modification. The specific post-translational modification may be the addition or deletion of a sugar chain. For example, the amino acid residue at position 297 (EU numbering) in the IgG1 constant region can be modified with a sugar chain. The structure of the sugar chain is not limited to a particular structure. For example, sialic acid can be added to the sugar chain of the Fc region (MAbs 2(5), September-October 2010, pp. 519-527). Typically, antibodies that are expressed in eukaryotic cells contain a glycosylation site in the constant region. For example, it is known that several types of sugar chains are normally added to antibodies expressed in cells, such as naturally occurring antibody-producing mammalian cells or eukaryotic cells transformed with an expression vector that contains the DNA encoding the antibody.

In the context of the present description, eukaryotic cells include yeast and animal cells. For example, CHO cells and HEK293 cells are representative animal cells that are used for transformation with an expression vector containing DNA that encodes an antibody. On the other hand, constant regions without glycosylation also fall within the scope of the present invention. Antibodies whose constant regions are not glycosylated can be generated by expression of an antibody-encoding gene in prokaryotic cells such as Escherichia coli.

In addition, the polypeptide containing the variant Fc region of the present invention can be chemically modified with various molecules such as polyethylene glycol (PEG) and cytotoxic substances. Methods for said chemical modification of polypeptides have been developed in the art.

One object of the invention is an isolated polypeptide containing a variant Fc region with increased Fc-gamma RIIb-binding activity. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In some aspects of the invention, the antibody is a chimeric antibody or a humanized antibody. The origin of the antibody is not limited to a particular origin, but examples include human antibody, mouse antibody, rat antibody, and rabbit antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein.

Antibody variable regions that contain the Fc region variant as described herein and protein-binding motifs of Fc-containing fusion proteins that contain the Fc region variant can recognize any antigen. Examples of antigens that can be bound by such antibodies and fusion proteins include, but are not limited to, ligands (cytokines, chemokines, etc.), receptors, cancer antigens, MHC antigens, differentiation antigens, immunoglobulins, and immune complexes, in part including immunoglobulins.

Examples of cytokines that can be bound by an antibody or fusion protein containing an Fc region variant of the invention and/or recombinantly fused to a polypeptide that contains said Fc region variant include, but are not limited to, interleukins 1-18 , colony stimulating factors (G-CSF, M-CSF, GM-CSF, etc.), interferons (IFN-alpha, IFN-beta, IFN-gamma, etc.), growth factors (EGF, FGF, IGF , NGF, PDGF, TGF, HGF, etc.), tumor necrosis factors (TNF-alpha and TNF-beta), lymphotoxin, erythropoietin, leptin, SCF, TPO, MCAF and BMP.

Examples of chemokines that can be bound by an antibody or fusion protein containing an Fc region variant of the invention and/or recombinantly fused to a polypeptide that contains said Fc region variant include, but are not limited to, CC chemokines, such as CCL1-CCL28, CXC chemokines such as CXCL1-CXCL17, C chemokines such as XCL1-XCL2, and CX3C chemokines such as CX3CL1.

Examples of receptors that can be bound by an antibody or fusion protein containing an Fc region variant of the invention and/or recombinantly fused to a polypeptide that contains said Fc region variant include, but are not limited to, receptors belonging to receptor families such as the hematopoietic growth factor receptor family, the cytokine receptor family, the tyrosine kinase-type receptor family, the serine/threonine kinase-type receptor family, the TNF receptor family, the G protein-coupled receptor family, the GPI-anchored receptor family, the tyrosine phosphatase-type receptor family , the adhesion factor family, and the hormone receptor family. The receptors belonging to these receptor families and their characteristics are described in many documents such as New Comprehesive Biochemistry ed. Cooke, vol. 18B "Hormones and their Actions Part II", 1988, pp. 1-46, Elsevier Science Publishers BV; Patthy, Cell 61(1), 1990, pp. 13-14; Ullrich, Cell 61(2), 1990, pp. 203-212; Massague, Cell 69(6), 1992, pp. 1067-1070; Miyajima et al., Annu. Rev. Immunol. 10, 1992, pp. 295-331; Taga et al., FASEB J. 6, 1992, pp. 3387-3396; Fantl et al., Annu. Rev. Biochem. 62, 1993, pp. 453-481; Smith et al., Cell 76(6), 1994, pp. 959-962 and Flower, Biochim. Biophys. Acta 1422(3), 1999, pp. 207-234.

и др., Cell 57 (2), 1989, cc. 277-285), человеческие или мышиные рецепторы гранулоцитарного колониестимулирующего фактора (G-CSF) (Fukunaga и др., Proc. Natl. Acad. Sci. USA 87 (22), 1990, cc. 8702-8706), mG-CSFR (Fukunaga и др., Cell 61(2), 1990, cc. 341-350), человеческие или мышиные рецепторы тромбопоэтина (ТРО) (Vigon и др., Proc. Natl. Acad. Sci. USA. 89 (12), 1992, cc. 5640-5644; Skoda и др., EMBO J. 12 (7), 1993, cc. 2645-2653), человеческие или мышиные рецепторы инсулина (Ullrich и др., Nature 313 (6005), 1985, cc. 756-761), человеческие или мышиные рецепторы для лиганда Flt-3 (Small и др., Proc. Natl. Acad. Sci. USA. 91 (2), 1994, cc. 459-463), человеческие или мышиные рецепторы тромбоцитарного фактора роста (PDGF) (Gronwald и др., Proc. Natl. Acad. Sci. USA. 85 (10), 1988, cc. 3435-3439), человеческие или мышиные рецепторы интерферона (IFN)-альфа и -бета (Uze и др., Cell 60 (2), 1990, cc. 225-234 (1990); Novick и др, Cell 77 (3), 1994, cc. 391-400), человеческие или мышиные рецепторы лептина, человеческие или мышиные рецепторы гормонов роста (GH), человеческие или мышиные рецепторы интерлейкина (IL)-10, человеческие или мышиные рецепторы инсулиноподобного фактора роста (IGF)-I, человеческие или мышиные рецепторы ингибирующего лейкоз фактора (LIF), и человеческие или мышиные рецепторы цилиарного нейротрофического фактора (CNTF).Examples of specific receptors belonging to the above receptor families include human or mouse erythropoietin (EPO) receptors (Jones et al., Blood 76 (1), 1990, pp. 31-35;

et al., Cell 57(2), 1989, pp. 277-285), human or mouse granulocyte colony stimulating factor (G-CSF) receptors (Fukunaga et al., Proc. Natl. Acad. Sci. USA 87 (22), 1990, pp. 8702-8706), mG-CSFR ( Fukunaga et al., Cell 61(2), 1990, pp. 341-350), human or murine thrombopoietin (TPO) receptors (Vigon et al., Proc. Natl. Acad. Sci. USA. 89 (12), 1992 , pp. 5640-5644; Skoda et al., EMBO J. 12 (7), 1993, pp. 2645-2653), human or murine insulin receptors (Ullrich et al., Nature 313 (6005), 1985, pp. 756-761), human or mouse receptors for Flt-3 ligand (Small et al., Proc. Natl. Acad. Sci. USA. 91 (2), 1994, pp. 459-463), human or mouse platelet factor receptors (PDGF) (Gronwald et al., Proc. Natl. Acad. Sci. USA. 85 (10), 1988, pp. 3435-3439), human or mouse interferon receptors (IFN)-alpha and -beta (Uze and et al., Cell 60 (2), 1990, pp. 225-234 (1990); Novick et al., Cell 77 (3), 1994, pp. 391-400), human or mouse leptin receptors, human or murine growth hormone (GH) receptors, human or murine interleukin (IL)-10 receptors, human or murine insulin-like growth factor (IGF)-I receptors, human or murine leukemia inhibitory factor (LIF) receptors, and human or murine ciliary receptors. neurotrophic factor (CNTF).

Cancer antigens are antigens that are expressed on cells that become cancerous and are also referred to as tumor-specific antigens. Abnormal sugar chains that appear on the cell surface or on protein molecules when cells become cancerous are also referred to as cancer antigens and are also called sugar chain cancer antigens. Examples of cancer antigens that can be bound by an antibody or fusion protein containing a variant Fc region of the invention include, but are not limited to, GPC3, which is a receptor belonging to the above family of GPI-anchored receptors and is also expressed in some cancers, including liver cancer (Midorikawa et al., Int. J. Cancer 103(4), 2003, pp. 455-465) and EpCAM, which is expressed in some cancers, including lung cancer (Linnenbach and et al., Proc. Natl, Acad. Sci. USA 86 (1), 1989, pp. 27-31), CA19-9, CA15-3 and sialyl-SSEA-1 (SLX).

MHC antigens are roughly classified into MHC class I antigens and MHC class II antigens. Class I MHC antigens include HLA-A, -B, -C, -E, -F, -G and -H, and class II MHC antigens include HLA-DR, -DQ and -DP.

Examples of differentiation antigens that can be bound by an antibody or fusion protein containing an Fc region variant of the invention and/or recombinantly fused to a polypeptide that contains said Fc region variant include, but are not limited to, CD1, CD2 , CD4, CD5, CD6, CD7, CD8, CD10, CD11a, CD11b, CD11c, CD13, CD14, CD15s, CD16, CD18, CD19, CD20, CD21, CD23, CD25, CD28, CD29, CD30, CD32, CD33, CD34 , CD35, CD38, CD40, CD41a, CD41b, CD42a, CD42b, CD43, CD44, CD45, CD45RO, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD51, CD54, CD55, CD56, CD57, CD58, CD61 , CD62E, CD62L, CD62P, CD64, CD69, CD71, CD73, CD95, CD102, CD106, CD122, CD126 and CDw130.

Immunoglobulins include IgA, IgM, IgD, IgG and IgE. The immune complexes include a component of at least any of the immunoglobulins.

Other examples of antigens that can be bound by an antibody or fusion protein containing an Fc region variant of the invention and/or recombinantly fused to a polypeptide that contains said Fc region variant include, but are not limited to, 17-IA , 4-1BB, 4Dc, 6-keto-PGF1a, 8-iso-PGF2a, 8-oxo-dG, A1 adenosine receptor, A33, ACE, ACE-2, activin, activin A, activin AB, activin B, activin C, activin RIA, activin RIA ALK-2, activin RIB ALK-4, activin RIIA, activin RIIB, ADAM, ADAM10, ADAM12, ADAM15, ADAM17/TACE, ADAM8, ADAM9, ADAMTS, ADAMTS4, ADAMTS5, addressin, aFGF, ALCAM , ALK, ALK-1, ALK-7, alpha-1 antitrypsin, alpha-V/beta antagonist, ANG, Ang, APAF-1, APE, APJ, APP, APRIL, AR, ARC, ART, artemin, anti- Id, ASPARTIC, atrial natriuretic peptide, av/b3-integrin, Ax1, b2M, B7-1, B7-2, B7-H, B-lymphocyte-stimulating factor (BlyS), BACE, BACE-1, Bad, BAFF, BAFF -R, Bag-1, BAK, Vah, BCA-1, BACAM, Bcl, BCMA, BDNF, b-ECGF, bFGF, BID, Bik, BIM, BLC, BL-CAM, BLK, BMP, BMP-2 BMP-2a, osteogenin BMP-3, BMP-4, BMP-2b, BMP-5, BMP-6 Vgr-1, BMP-7 (OP-1), BMP-8 (BMP- 8a, OP-2), BMPR, BMPR-IA (ALK-3), BMPR-IB (ALK-6), BRK-2, RPK-1, BMPR-II (BRK-3), BMP, b-NGF, BOK, bombesin, bone neurotrophic factor, BPDE, BPDE-DNA, BTC, complement system factor 3 (C3), C3a, C4, C5, C5a, C10, CA125, CAD-8, calcitonin, cAMP, carcinoembryonic antigen (CEA), cancer-associated antigen, cathepsin A, cathepsin B, cathepsin C/DPPI, cathepsin D, cathepsin E, cathepsin H, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin X/Z/P, CBL, CCI, CCK2, CCL, CCL1, CCL11, CCL12, CCL13, CCL14, CCL15, CCL16, CCL17, CCL18, CCL19, CCL2, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CCL28, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9/10, CCR, CCR1, CCR10, CCR11, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD1, CD2, CD3, CD3E, CD4, CD5, CD6, CD7, CD8, CD10, CD11a, CD11b, CD11c, CD13, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD27L, CD28, CD29, CD 30, CD30L, CD32, CD33 (p67 protein), CD34, CD38, CD40, CD40L, CD44, CD45, CD46, CD49a, CD52, CD54, CD55, CD56, CD61, CD64, CD66e, CD74, CD80 (B7-1) , CD89, CD95, CD123, CD137, CD138, CD140a, CD146, CD147, CD148, CD152, CD164, CEACAM5, CFTR, cGMP, CINC, botulinum toxin, Clostridium perfringens toxin, CKb8-1, CLC, CMV, CMV UL, CNTF , CNTN-1, COX, C-Ret, CRG-2, CT-1, CTACK, CTGF, CTLA-4, CX3CL1, CX3CR1, CXCL, CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9 , CXCL10, CXCL11, CXCL12, CXCL13, CXCL14, CXCL15, CXCL16, CXCR, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, tumor cytokeratin-associated antigen, DAN, DCC, DcR3, DC-SIGN, complement regulating factor ( dissociation accelerator factor), des(1-3)-IGF-I (IGF-1 brain), Dhh, digoxin, DNAM-1, DKase, Dpp, DPPIV/CD26, Dtk, ECAD, EDA, EDA-A1, EDA -A2, EDAR, EGF, EGFR (ErbB-1), EMA, EMMPRIN, ENA, endothelin receptor, enkephalinase, eNOS, Eot, eotaxin 1, EpCAM, ephrin B2/EphB4, EPO, ERCC, E-selectin, ET-1 , factor IIa , factor VII, factor VIIIc, factor IX, fibroblast-activating protein (FAP), Fas, FcR1, FEN-1, ferritin, FGF, FGF-19, FGF-2, FGF3, FGF-8, FGFR, FGFR-3, fibrin, FL, FLIP, Flt-3, Flt-4, follicle-stimulating hormone, fractalkin, FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, FZD10, G250, Gas6, GCP-2, GCSF, GD2 , GD3, GDF, GDF-1, GDF-3 (Vgr-2), GDF-5 (BMP-14, CDMP-1), GDF-6 (BMP-13, CDMP-2), GDF-7 (BMP- 12, CDMP-3), GDF8 (myostatin), GDF-9, GDF-15 (MIC-1), GDNF, GFAP, GFRa-1, GFR-alpha1, GFR-alpha2, GFR-alpha3, GITR, glucagon, Glut4 , glycoprotein IIb/IIIa (GPIIb/IIIa), GM-CSF, gp130, gp72, GRO, growth hormone releasing factor, hapten (NP-cap or NIP-cap), HB-EGF, HCC, HCMV envelope glycoprotein gB, envelope HCMV gH glycoprotein, HCMV UL, hematopoietic growth factor (HGF), Her B gp120, heparanase, Her2, Her2/neu (ErbB-2), Her3 (ErbB-3), Her4 (ErbB-4), herpes virus envelope glycoprotein gB simple (HSV), glycoprotein HSV gD, HGFA, high molecular weight associated with me lanoma antigen (HMW-MAA), HIV gp120, HIV IIIB gp 120 V3-loop, HLA, HLA-DR, HM1.24, HMFG PEM, HRG, Hrk, human cardiac myosin, human cytomegalovirus (HCMV), human growth hormone ( HGH), HVEM, I-309, IAP, ICAM, ICAM-1, ICAM-3, ICE, ICOS, IFNg, Ig, IgA receptor, IgE, IGF, IGF binding protein, IGF-1R, IGFBP, IGF- I, IGF-II, IL, IL-1, IL-1R, IL-2, IL-2R, IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL- 8, IL-9, IL-10, IL-12, IL-13, IL-15, IL-18, IL-18R, IL-23, interferon (IFN)-alpha, IFN-beta, IFN-gamma, inhibin , iNOS, insulin A chain, insulin B chain, insulin-like growth factor1, integrin alpha2, integrin alpha3, integrin alpha4, integrin alpha4/beta1, integrin alpha4/beta7, integrin alpha5 (alpha V), integrin alpha5/beta1, integrin alpha5/beta3 , integrin alpha6, integrin beta1, integrin beta2, interferon gamma, IP-10, I-TAC, JE, kallikrein 2, kallikrein 5, kallikrein 6, kallikrein 11, kallikrein 12, kallikrein 14, kallikrein 15, kallikrein L1, kallikrein L2, kallikrein L 3, kallikrein L4, KC, KDR, keratinocyte growth factor (KGF), laminin 5, LAMP, LAP, LAP (TGF-1), latent TGF-1, latent TGF-1 bp1, LBP, LDGF, LECT2, Lefty factor, Lewis-Y antigen, Lewis-associated antigen, LFA-1, LFA-3, Lfo, LIF, LIGHT, lipoprotein, LIX, LKN, Lptn, L-selectin, LT-a, LT-b, LTB4, LTBP- 1, lung surface antigen, luteinizing hormone, lymphotoxin receptor beta, Mac-1, MAdCAM, MAG, MAP2, MARC, MCAM, MSK-2, MCP, M-CSF, MDC, Mer, metalloprotease, MGDF receptor, MGMT, MHC (HLA-DR), MIF, MIG, MIP, MIP-1-alpha, MK, MMAC1, MMP, MMP-1, MMP-10, MMP-11, MMP-12, MMP-13, MMP-14, MMP- 15, MMP-2, MMP-24, MMP-3, MMP-7, MMP-8, MMP-9, MPIF, Mpo, MSK, MSP, Mucin (Muc1), MUC18, Müller inhibitory substance, Mug, MuSK, NAIP , NAP, NCAD, N-cadherin, NCA 90, NCAM, neprilysin, neurotrophin-3, -4 or -6, neuroturin, nerve growth factor (NGF), NGFR, NGF-beta, nNOS, NO, NOS, Npn, NRG -3, NT, NTN, OB, OGG1, OPG, OPN, OSM, OX40L, OX40R, p150, p95, PADPr, parathyroid hormone, PARC, PARP, PBR, PBSF, PCAD, P-cadherin, PCNA, PDGF, PDK-1, PECAM, PEM, PF4, PGE, PGF, PGI2, PGJ2, PIN, PLA2, placental alkaline phosphatase (PLAP), PIGF, PLP , PP14, proinsulin, prorelaxin, protein C, PS, PSA, PSCA, prostate-specific membrane antigen (PSMA), PTEN, PTHrp, Ptk, PTN, R51, RANK, RANKL, RANTES, relaxin A chain, relaxin B chain, renin, respiratory -syncytial virus (RSV) F, RSV Fgp, Ret, rheumatoid factor, RLIP76, RPA2, RSK, S100, SCF/KL, SDF-1, serine, serum albumin, sFRP-3, Shh, SIGIRR, SK-1, SLAM , SLPI, SMAC, SMDF, SMOH, SOD, SPARC, Stat, STEAP, STEAP-II, TACE, TACI, TAG-72 (tumor-associated glycoprotein-72), TARC, TCA-3, T cell receptor (eg, T cell receptor alpha/beta), TdT, TESK, TEM1, TEM5, TEM7, TEM8, TERT, PLAP-like testicular alkaline phosphatase, TfR, TGF, TGF-alpha, TGF-beta, TGF-beta Pan-specific, TGF -betaRI (ALK-5), TGF-betaRII, TGF-betaRIIb, TGF-betaRIII, TGF-beta 1, TGF-beta2, TGF-beta3, TGF-beta4, TGF-beta5, tro mbin, thymus Ck-1, thyroid-stimulating hormone, Tie, TIMP, TIQ, tissue factor, TMEFF2, Tmpo, TMPRSS2, TNF, TNF-alpha, TNF-alpha, beta, TNF-beta2, TNFc, TNF-RI, TNF-RII , TNFRSF10A (TRAIL R1 Apo-2, DR4), TNFRSF10B (TRAIL R2 DR5, KILLER, TRICK-2A, TRICK-B), TNFRSF10C (TRAIL R3 DcR1, LIT, TRID), TNFRSF10D (TRAIL R4 DcR2, TRUNDD), TNFRSF11A (RANK ODF R, TRANCE R), TNFRSF11B (OPG OCIF, TR1), TNFRSF12 (TWEAK R FN14), TNFRSF13B (TACI), TNFRSF13C (BAFF R), TNFRSF14 (HVEM ATAR, HveA, LIGHT R, TR2), TNFRSF16 ( NGFR p75NTR), TNFRSF17 (BCMA), TNFRSF18 (GITR AITR), TNFRSF19 (TROY TAJ, TRADE), TNFRSF19L (RELT), TNFRSF1A (TNF RI CD120a, p55-60), TNFRSF1B (TNF R00 CD 120b, p75-80) , TNFRSF26 (TNFRH3), TNFRSF3 (LTbR TNF RIII, TNFC R), TNFRSF4 (OX40 ACT35, TXGP1 R), TNFRSF5 (CD40 p50), TNFRSF6 (Fas Apo-1, APT1, CD95), TNFRSF6B (DcR3 M68, TR6) , TNFRSF7 (CD27), TNFRSF8 (CD30), TNFRSF9 (4-1BB CD 137, ILA), TNFRSF21 (DR6), TNFRSF22 (DcTRAIL R2 TNFRH2), TNFRST23 (DcTRAIL R1 TNFRH1), TNFRSF25 (DR3 Apo-3, LARD, TR-3, T RAMP, WSL-1), TNFSF10 (TRAIL Apo-2 ligand, TL2), TNFSF11 (TRANCE/RANK ODF ligand, OPG ligand), TNFSF12 (TWEAK Apo-3 ligand, DR ligand), TNFSF13 (APRIL TALL2), TNFSF13B (BAFF BLYS, TALL1, THANK, TNFSF20), TNFSF14 (LIGHT HVEM ligand, LTg), TNFSF15 (TL1A/VEGI), TNFSF18 (GITR ligand AITR ligand, TL6), TNFSF1A (TNF-a conectin , DIF, TNFSF2), TNFSF1B (TNF-b LTa, TNFSF1), TNFSF3 (LTb TNFC, p33), TNFSF4 (OX40 ligand gp34, TXGP1), TNFSF5 (CO40 ligand CD154, gP39, HIGM1, IMD3, TRAP), TNFSF6 (Fas ligand Apo-1 ligand, APT1 ligand), TNFSF7 (CD27 ligand CD70), TNFSF8 (CD30 ligand CD153), TNFSF9 (4-1BB ligand CD137 ligand), TP-1, t- PA, Tpo, TRAIL, TRAIL R, TRAIL-R1, TRAIL-R2, TRANCE, transferrin receptor, TRF, Trk, TROP-2, TSG, TSLP, tumor-associated antigen CA125, tumor-associated antigen expressing associated Lewis antigen -Y carbohydrates, TWEAK, TXB2, Ung, uPAR, uPAR-1, urokinase, VCAM, VCAM-1, VECAD, VE-cadherin, VE-cadherin-2, VEFGR-1 (flt-1), VEGF, VEGFR, VEGFR -3 (flt-4), VEGI, VIM, viral antigen, VLA, VLA-1, VLA-4, VNR integrin, von Willebrand factor, WIF-1, WNT1, WNT2, WNT2B/13, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, WNT16, XCL1, XCL2, XCR1, XCR1, XEDAR, XIAP, XPD, HMGB1, IgA, A beta, CD81, CD97, CD98 , DDR1, DKK1, EREG, Hsp90, IL-17/IL-17R, IL-20/IL-20R, oxidized LDL, PCSK9, prekallikrein, RON, TMEM16F, SOD1, chromogranin A, chromogranin B, tau, VAP1, high molecular weight kininogen , IL-31, IL-31R, Nav1.1, Nav1.2, Nav1.3, Nav1.4, Nav1.5, Nav1.6, Nav1.7, Nav1.8, Nav1.9, EPCR, C1, C1q , C1r, C1s, C2, C2a, C2b, C3, C3a, C3b, C4, C4a, C4b, C5, C5a, C5b, C6, C7, C8, C9, factor B, factor D, factor H, properdin, sclerostin, fibrinogen, fibrin, prothrombin, thrombin, tissue factor, factor V, factor Va, factor VII, factor VIIa, factor VIII, factor VIIIa, factor IX, factor IXa, factor X, factor Xa, factor XI, factor XIa, factor XII, factor XIIa, factor XIII, factor XIIIa, TFPI, antithrombin III, EPCR, thrombomodulin, TAPI, tPA, plasminogen, plasmin, PAI-1, PAI-2, GPC3, syndecan-1, syndecan-2, syndecan-3, syndecan-4, LPA and S1P; and receptors for hormones and growth factors.

As stated in the present description, the presence of changes in one or more amino acid residues in amino acid sequences that form variable regions is acceptable if their antigen-binding activity is maintained. Changes affecting the amino acid sequence of the variable region are not limited to the site at which the change is made and the number of amino acids changed. For example, the amino acids present in the CDR and/or FR can be modified accordingly. When the amino acids in the variable region are changed, the binding activity is preferably maintained without particular limitation; and, for example, compared to the activity before the change, the binding activity can be 50% or more, 80% or more, and 100% or more. In addition, binding activity can be increased due to amino acid changes. For example, the binding activity may be 2-fold, 5-fold, 10-fold, or the like. compared to activity before the change. The amino acid sequence change may be a substitution, addition, deletion or modification of at least one amino acid residue.

For example, modification of the N-terminal glutamine of the variable region, resulting in the formation of pyroglutamic acid by pyroglutamylation, is well known to those skilled in the art. Thus, when the N-terminus of the heavy chain is glutamine, the antibodies described herein may contain variable regions in which glutamine is modified to form pyroglutamic acid.

The variable regions of the antibodies described herein may be of any sequence and may be the variable regions of antibodies of any origin, e.g. mouse antibodies, rat antibodies, rabbit antibodies, goat antibodies, camel antibodies, humanized antibodies obtained by humanizing said non-human antibodies, and human antibodies. In addition, these antibodies may have various amino acid substitutions introduced into their variable regions to improve their antigen binding, pharmacokinetic characteristics, stability, and immunogenicity. Variable regions may have the ability to re-bind to antigens due to their pH dependent antigen binding (WO 2009/125825).

In the constant regions of the light chains of antibodies, the kappa chain and the lambda chain are present, either of which is acceptable. In addition, they may have amino acid changes such as substitutions, deletions, additions and/or insertions.

In addition, polypeptides that contain variants of the Fc regions described herein can form Fc-containing fusion proteins by binding to other proteins, such as physiologically active peptides. Said fusion proteins may be multimers of at least two polypeptides containing variants of the Fc regions. Examples of other proteins are (but are not limited to) receptors, adhesion molecules, ligands, and enzymes.

Examples of Fc-containing fusion proteins include proteins fused via an Fc region to a receptor that binds to a target molecule, including TNFR-Fc fusion protein, IL1R-Fc fusion protein, VEGFR-Fc fusion protein, and CTLA4-Fc fusion protein (Economides Nat Med 9(1) 2003 47-52 Dumont et al BioDrugs 20(3) 2006 1 51-160). In addition, the proteins to be fused may be other molecules that have the ability to bind to the target, such as scFv (WO 2005/037989), single domain antibodies (WO 2004/058821; WO 2003/002609), antibody-like molecules (Davinder, Curr Op Biotech 17 2006 pp 653-658 Current Opinion in Biotechnology 18 2007 pp 1-10 Nygren et al Curr op Struct Biol 7 1997 463 -469 and Hoss Protein Science 15, 2006, pp. 14-27 (2006), such as DARPin (WO 2002/020565), Affibody (WO 1995/001937), Avimer (WO 2004/044011 ; WO 2005/040229) and Adnectin (WO 2002/032925).Furthermore, antibodies and Fc-containing fusion proteins can be multispecific and can bind to several types of target molecules or epitopes.

B. Methods of recombination and composition

Antibodies can be obtained using recombination and composition techniques such as those described in US No. 4,816,567. One embodiment of the invention is an isolated nucleic acid encoding an anti-myostatin antibody as described herein. Another embodiment of the invention is an isolated nucleic acid that encodes a polypeptide containing a variant Fc region or parental Fc region as defined herein. Said nucleic acid may encode an amino acid sequence containing the VL and/or an amino acid sequence containing the VH of an antibody (eg, antibody light and/or heavy chains). The following embodiment of the invention is(are) one or more vectors (eg, expression vectors), which(e) contains(at) the specified nucleic acid. Another embodiment of the invention is a host cell that contains the specified nucleic acid. In one of these embodiments, the host cell contains (for example, as a result of transformation with these vectors): (1) a vector containing a nucleic acid that encodes an amino acid sequence containing the VL of the antibody, and an amino acid sequence containing the VH of the antibody, or (2 ) a first vector containing a nucleic acid that encodes an amino acid sequence containing the VL of an antibody, and a second vector containing a nucleic acid that encodes an amino acid sequence containing the VH of an antibody. In one embodiment, the host cell is a eukaryotic cell, such as a Chinese Hamster Ovary (CHO) cell or a lymphoid cell (eg, Y0-, NS0-, Sp20 cell). One embodiment of the invention is a method for producing an anti-myostatin antibody comprising culturing a host cell containing a nucleic acid that encodes the antibody described above under conditions suitable for expressing the antibody, and optionally isolating the antibody from the host cell (or culture medium). host cells). Another embodiment of the invention is a method for producing a polypeptide that contains a variant Fc region or a parent Fc region, comprising culturing a host cell that contains nucleic acid(s) encoding(s) a polypeptide, such as an antibody, Fc- an Fc region or variant as defined above under conditions suitable for expression of the polypeptide, and optionally isolating the polypeptide from the host cell (or host cell culture medium).

For recombinant production of an anti-myostatin antibody, antibody-encoding nucleic acids, such as those described above, are isolated and inserted into one or more vectors for further cloning and/or expression in a host cell. For recombinant production of an Fc region, a nucleic acid encoding an Fc region is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell. The specified nucleic acid can be easily isolated and sequenced using conventional procedures (for example, by using oligonucleotide probes that have the ability to specifically bind to the genes encoding the heavy and light chains of antibodies).

Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells as described herein. For example, antibodies can be made in bacteria, particularly when glycosylation and Fc-related effector function are not required. For expression of antibody fragments and polypeptides in bacteria, see, for example, US Nos. 5,648,237, 5,789,199, and 5,840,523 (see also Charlton in: Methods in Molecular Biology, ed. Lo B.K.C, Humana Press , Totowa, NJ, Vol.

In addition to prokaryotic organisms, eukaryotic microorganisms such as filamentous fungi or yeasts, including strains of fungi and yeasts whose glycosylation pathways have been "humanized" can be used as hosts suitable for cloning or expressing plasmids that encode antibodies, which makes it possible to obtain an antibody with partially or a fully human glycosylation scheme (see Gerngross, Nat. Biotech. 22, 2004, pp. 1409-1414 and Li et al., Nat. Biotech. 24, 2006, pp. 210-215).

Host cells that can be used to express the glycosylated antibody are also obtained from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells are insect cells, and plant cells can also be used. Numerous strains of baculoviruses and corresponding suitable insect cells as hosts have been identified, primarily for transfection of Spodoptera frugiperda cells.

Plant cell cultures can also be used as hosts (see, for example, US Nos. 5959177, 6040498, 6420548, 7125978 and 6417429 (description of PLANTIBODIES™ technology for producing antibodies in transgenic plants).

Vertebrate cells can also be used as hosts. For example, mammalian cell lines that are adapted to growth in suspension can be used. Other examples of suitable mammalian host cell lines are the CV1 monkey kidney cell line transformed with OB40 (COS-7); a human embryonic kidney cell line (HEK 293 cells or cell line 293, described, for example, in Graham et al., J. Gen. Virol., 36, 1977, pp. 59-74); baby hamster kidney cells (BHK); mouse Sertoli cells (TM4 cells described, for example, in Mather, Biol. Reprod., 23, 1980, pp. 243-251); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma (HELA) cells; dog kidney cells (MDCK); bovine rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor cells (MMT 060562); TRI cells as described, for example, in Mather et al., Annals NY Acad. Sci., 383, 1982, pp. 44-68); MRC 5 cells and FS4 cells. Other valuable mammalian host cell lines are Chinese hamster ovary (CHO) cells, including DHFR ^-- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA, 77, 1980, pp. 4 216-4220) ; and myeloma cell lines such as Y0, NS0 and Sp2/0. For a review of specific mammalian host cell lines that can be used to produce antibodies, see, for example, Yazaki and Wu, in: Methods in Molecular Biology, ed. V.K.S. Lo, Humana Press, Totowa, NJ, vol. 248, 2003, pp. 255-268.

Antibodies with pH dependent characteristics can be obtained using screening and/or mutagenesis methods, such as those described in WO 2009/125825. Screening methods may include any process that identifies an antibody with pH-dependent binding characteristics in a population of antibodies specific for a particular antigen. In some embodiments, screening methods may involve measuring one or more binding parameters (eg, KD or kd) of individual antibodies in an initial antibody population, both at acidic and neutral pH. Antibody binding parameters can be measured, for example, using surface plasmon resonance or any other analytical method that allows you to quantitatively or qualitatively evaluate the binding characteristics of an antibody to a particular antigen. In some embodiments, screening methods may involve identifying an antibody whose binding to an antigen is characterized by an acid/neutral pH KD ratio of 2 or more. In other embodiments, screening methods may include identifying an antibody whose binding to antigen has a KD ratio at pH 5.8/pH 7.4 of 2 or more. Alternatively, screening methods may involve identifying an antibody whose binding to an antigen is characterized by an acid/neutral pH kd ratio of 2 or more. In other embodiments, screening methods may include identifying an antibody whose binding to an antigen has a pH 5.8/pH 7.4 kd ratio of 2 or more.

In another embodiment of the invention, mutagenesis methods may involve the deletion, substitution or addition of an amino acid in the heavy and/or light chain of an antibody to enhance the pH-dependent binding of the antibody to the antigen. In some embodiments, one or more variable domains of an antibody, such as one or more HVRs (eg, CDRs), can be mutated. For example, mutagenesis can result in the replacement of an amino acid in one or more HVRs (eg, CDRs) of an antibody with a different amino acid. In some embodiments of the invention, mutagenesis can result in the replacement of an amino acid in at least one HVR (eg, CDR) of the antibody with histidine. In some embodiments, "increased pH dependent binding" means that the mutant version of the antibody has a higher acid/neutral pH KD ratio, or a higher acid/neutral pH KD ratio, than the original "parent" (i.e., less pH dependent) version of the antibody prior to mutagenesis. In some embodiments of the invention, the mutant version of the antibody is characterized by a KD ratio at acidic/neutral pH of 2 or more. In some embodiments, the mutant version of the antibody has a KD ratio at pH 5.8/pH 7.4 of 2 or more. Alternatively, the mutant version of the antibody has an acid/neutral pH kd ratio of 2 or more. In some embodiments of the invention, the mutant version of the antibody is characterized by a kd ratio at pH 5.8/pH 7.4 of 2 or more.

Polyclonal antibodies are preferably produced in animals after several subcutaneous (sc) or intraperitoneal (ip) injections of the relevant antibody and adjuvant. The relevant antigen can be conjugated to a protein immunogenic in the species to be immunized, e.g., keyhole hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor, using a bifunctional or derivatizing agent, e.g., maleimidobenzoyl sulfosuccinimide ester (conjugation via cysteine residues), N -hydroxysuccinimide (conjugation via lysine residues), glutaraldehyde, succinic anhydride, SOCl ₂ or R ¹ N=C=NR, where R and R ¹ represent different alkyl groups.

Animals (typically non-human mammals) are immunized with the antigen, immunogenic conjugates or derivatives by combining, for example, 100 or 5 μg of protein or conjugate (for rabbits and mice, respectively) with 3 volumes of complete Freund's adjuvant, and the solution is administered intradermally into several places. After 1 month, the animals are booster vaccinated (revaccinated) using from 1/5 to 1/10 of the original amount of the peptide or conjugate in complete Freund's adjuvant by subcutaneous injection at several sites. After 7-14 days, blood samples of the animals are obtained and the serum antibody titer is assessed. Animals are revaccinated until the titer reaches a plateau. Preferably, the animals are revaccinated using a conjugate containing the same antigen but conjugated to a different protein and/or via a different cross-linking agent. Conjugates can also be obtained in recombinant cell culture as protein fusions. In addition, aggregating agents such as alum can be used to enhance the immune response.

Monoclonal antibodies are derived from a population of substantially homogeneous antibodies, ie. the individual antibodies in a population are identical except for possible naturally occurring mutations and/or post-translational modifications (eg, isomerization, amidation), which may be present in minor amounts. The adjective "monoclonal" indicates that the antibody is not present in a mixture of different antibodies.

For example, monoclonal antibodies can be obtained using the hybridoma method first described by Kohler et al., Nature 256 (5517), 1975, cc. 495-497. In the hybrid method, a mouse or other suitable host animal such as a hamster is immunized according to the method described above to produce lymphocytes that produce or are capable of producing antibodies that specifically bind to the protein used for immunization. Alternatively, lymphocytes can be immunized in vitro.

The immunizing agent will typically include an antigenic protein or a fusion variant thereof. Typically, either peripheral blood lymphocytes (PBL) are used if cells of human origin are required, or splenic or lymph node cells are used if non-human mammalian sources are used. The lymphocytes are then fused to an immortalized cell line using a suitable fusion agent such as polyethylene glycol to form a hybridoma cell (Goding, Monoclonal Antibodies: Principles and Practice, Academic Press, 1986, pp. 59-103).

Immortalized cell lines are generally transformed mammalian cells, primarily rodent, bovine and human myeloma cells. Typically, rat or mouse myeloma cell lines are used. The hybridoma cells thus obtained are seeded and grown in a suitable culture medium, which preferably contains one or more substances that inhibit the growth or survival of unfused parental myeloma cells. For example, if the parent myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), then the culture medium for hybridomas typically includes hypoxanthine, aminopterine, and thymidine (HAT medium), i.e. substances that interfere with the growth of HGPRT-deficient cells.

Preferred immortalized myeloma cells are cells that can be efficiently fused, that maintain a stable high level of antibody production by selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these, murine myeloma lines such as those derived from MOPC-21 and MPC-11 murine tumors, which are available from the Salk Institute Cell Distribution Center, San Diego, PA, are preferred. California, USA, and SP-2 cells (and their derivatives, eg X63-Ag8-653), which can be obtained from the American Type Culture Collection, Manassas, pc. Virginia, USA. Also described are human myeloma and mouse-human heteromyeloma cell lines that can be used for the production of human monoclonal antibodies (Kozbor et al., J. Immunol. 133 (6), 1984, cc. 3001-3005; Brodeur et al., Monoclonal Antibody Production Techniques and Applications, Marcel Dekker, Inc., New York, 1987, pp. 51-63).

The culture medium in which the hybridoma cells are grown is evaluated for the production of monoclonal antibodies against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by hybridoma cells is determined by immunoprecipitation or an in vitro binding assay such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). These techniques and assays are known in the art. For example, binding affinity can be determined using the Scatchard analysis described in Munson, Anal. Biochem. 107(1), 1980, pp. 220-239.

Once hybridoma cells are found to produce antibodies of the desired specificity, affinity and/or activity, clones can be subcloned using serial dilution procedures and grown using standard methods (Goding, supra). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. In addition, hybridoma cells can be grown in vivo as tumors in mammals.

Monoclonal antibodies secreted by subclones can be separated from the culture medium, ascitic fluid or serum using conventional immunoglobulin purification procedures, such as chromatography for protein A-sepharose, hydroxyapatite, gel electrophoresis, dialysis or affinity chromatography.

Antibodies can be obtained by immunizing a suitable host animal with an antigen. In one embodiment, the antigen is a full-length myostatin-containing polypeptide. In one of the embodiments of the invention, the antigen is a polypeptide containing latent myostatin. In one embodiment, the antigen is a polypeptide containing a myostatin propeptide. In one embodiment, the antigen is a polypeptide containing a region corresponding to amino acids at positions 21-100 of the myostatin propeptide (SEQ ID NO: 78). In one embodiment, the antigen is a polypeptide containing amino acids at positions 21-80, 41-100, 21-60, 41-80, 61-100, 21-40, 41-60, 61-80 or 81-100 myostatin propeptide (SEQ ID NO: 78). The present invention also relates to antibodies obtained by immunizing an animal with an antigen. Antibodies may have singly or in combination any of the features described above in the "Examples of Myostatin Antibodies" section.

The Fc region can be obtained by re-eluting the protein A adsorbed fraction after partial digestion of monoclonal antibodies as IgG1, IgG2, IgG3, IgG4 or the like. with a protease such as pepsin. A protease is not limited to a particular protease as long as it can cleave a full-length antibody to form Fab and F(ab') ₂ in a limited manner under appropriate enzymatic reaction conditions such as pH, and examples include pepsin and papain.

In addition, the present invention provides a method for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity compared to a polypeptide containing a parent Fc region, which comprises introducing at least one amino acid change into the parent Fc region. . In some aspects of the invention, the resulting polypeptide is an antibody. In some embodiments, the antibody is a chimeric antibody or a humanized antibody. In some aspects of the invention, the resulting polypeptide is an Fc-containing fusion protein.

In some embodiments of the invention, the method of obtaining is that the following steps are carried out: (a) a polypeptide containing the parent Fc region is obtained; (b) introducing at least one amino acid change into the parent Fc region in the polypeptide; (c) measuring the binding activity to simian Fc-gamma RIIb of the polypeptide that contains the parental Fc region and the polypeptide that contains the Fc region obtained in step (b); and (d) selecting a polypeptide containing a variant Fc region with increased binding activity to simian Fc-gamma RIIb compared to a polypeptide containing the parental Fc region.

In some embodiments of the invention, the method of obtaining includes the following stages, which: (a) receive a polypeptide containing the parental Fc region; (b) introducing at least one amino acid change into the parent Fc region in the polypeptide; (c) simian Fc-gamma RIIIa binding activity of the polypeptide that contains the parental Fc region and the polypeptide that contains the Fc region obtained in step (b) is measured; and (d) selecting a polypeptide containing a variant Fc region with reduced simian Fc-gamma RIIIa binding activity compared to a polypeptide containing the parental Fc region.

In some embodiments of the invention, the method of obtaining is that the following steps are carried out: (a) a polypeptide containing the parent Fc region is obtained; (b) introducing at least one amino acid change into the parent Fc region in the polypeptide; (c) measure the activity of binding to human Fc-gamma RIIb polypeptide, which contains the parental Fc region, and the polypeptide, which contains the Fc region obtained in stage (b); and (d) selecting a polypeptide containing a variant Fc region with increased binding activity to the human Fc-gamma RIIb compared to a polypeptide containing the parental Fc region.

In some embodiments of the invention, the method of obtaining includes the following stages, which: (a) receive a polypeptide containing the parental Fc region; (b) introducing at least one amino acid change into the parent Fc region in the polypeptide; (c) measuring the binding activity to human Fc-gamma RIIIa of the polypeptide that contains the parental Fc region and the polypeptide that contains the Fc region obtained in step (b); and (d) selecting a polypeptide containing a variant Fc region with reduced binding activity to the human Fc gamma RIIIa compared to a polypeptide containing the parental Fc region.

In some embodiments of the invention, the method of obtaining is that the following steps are carried out: (a) a polypeptide containing the parent Fc region is obtained; (b) introducing at least one amino acid change into the parent Fc region in the polypeptide; (c) measuring the activity of binding to human Fc-gamma RIIa (H-type) of the polypeptide that contains the parental Fc region and the polypeptide that contains the Fc region obtained in step (b); and (d) selecting a polypeptide containing a variant Fc region with reduced binding activity to the human Fc gamma RIIa (H-type) compared to a polypeptide containing the parental Fc region.

In some embodiments of the invention, the method of obtaining is that the following steps are carried out: (a) a polypeptide containing the parent Fc region is obtained; (b) introducing at least one amino acid change into the parent Fc region in the polypeptide; (c) measuring the activity of binding to human Fc-gamma RIIa (R-type) of the polypeptide that contains the parental Fc region and the polypeptide that contains the Fc region obtained in step (b); and (d) selecting a polypeptide containing a variant Fc region with reduced binding activity to the human Fc gamma RIIa (R-type) compared to a polypeptide containing the parental Fc region.

In one aspect of the invention, at least one amino acid is altered in accordance with the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity at at least one position selected from the group consisting of: 231 , 232, 233, 234, 235, 236, 237, 238, 239, 264, 266, 267, 268, 271, 295, 298, 325, 326, 327, 328, 330, 331, 332, 334 and 396, according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In another aspect of the invention, one amino acid is changed in accordance with the method described above for obtaining a polypeptide containing a variant Fc region with increased Fc-gamma RIIb-binding activity, at position 236.

In another aspect of the invention, at least two amino acids are changed in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity, where the changes are: (a) one amino acid change at position 236 and ( b) at least one amino acid change in at least one position selected from the group consisting of: 231, 232, 233, 234, 235, 237, 238, 239, 264, 266, 267, 268, 271, 295, 298, 325, 326, 327, 328, 330, 331, 332, 334 and 396 according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In another aspect of the invention, at least two amino acids are changed in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity, where the changes are: (a) one amino acid change at position 236, and (b) at least one amino acid change at at least one position selected from the group consisting of: 231, 232, 235, 239, 268, 295, 298, 326, 330 and 396, according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In another aspect of the invention, at least two amino acids are changed in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity, where the changes are: (a) one amino acid change at position 236 and ( b) at least one amino acid change in at least one position selected from the group consisting of: 268, 295, 326, and 330, according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In another aspect of the invention, the amino acids are changed in accordance with the method described above for obtaining a polypeptide containing an Fc region variant with increased Fc-gamma RIIb-binding activity, at one of the following positions (1)-(37): (1) at positions 231, 236, 239, 268 and 330; (2) in regulations 231, 236, 239, 268, 295 and 330; (3) in regulations 231, 236, 268 and 330; (4) in regulations 231, 236, 268, 295 and 330; (5) in regulations 232, 236, 239, 268, 295 and 330; (6) in regulations 232, 236, 268, 295 and 330; (7) in regulations 232, 236, 268 and 330; (8) in regulations 235, 236, 268, 295, 326 and 330; (9) in regulations 235, 236, 268, 295 and 330; (10) in regulations 235, 236, 268 and 330; (11) in regulations 235, 236, 268, 330 and 396; (12) in regulations 235, 236, 268 and 396; (13) in regulations 236, 239, 268, 295, 298 and 330; (14) in regulations 236, 239, 268, 295, 326 and 330; (15) in regulations 236, 239, 268, 295 and 330; (16) in regulations 236, 239, 268, 298 and 330; (17) in regulations 236, 239, 268, 326 and 330; (18) in regulations 236, 239, 268 and 330; (19) in regulations 236, 239, 268, 330 and 396; (20) in regulations 236, 239, 268 and 396; (21) in regulations 236 and 268; (22) in regulations 236, 268 and 295; (23) in regulations 236, 268, 295, 298 and 330; (24) in regulations 236, 268, 295, 326 and 330; (25) in regulations 236, 268, 295, 326, 330 and 396; (26) in regulations 236, 268, 295 and 330; (27) in regulations 236, 268, 295, 330 and 396; (28) in regulations 236, 268, 298 and 330; (29) in regulations 236, 268, 298 and 396; (30) in regulations 236, 268, 326 and 330; (31) in regulations 236, 268, 326, 330 and 396; (32) in regulations 236, 268 and 330; (33) in regulations 236, 268, 330 and 396; (34) in regulations 236, 268 and 396; (35) in regulations 236 and 295; (36) in regulations 236, 330 and 396; and (37) in provisions 236 and 396 according to the EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In a further aspect of the invention, an amino acid change in accordance with the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity is selected at each of positions selected from the group consisting of: (a) Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, Tyr at position 231; (6) Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, Tyr at position 232; (c) Asp at position 233; (d) Trp, Tyr at position 234; (e) Trp at position 235; (f) Ala, Asp, Glu, His, Ile, Leu, Met, Asn, Gln, Ser, Thr, Val at position 236; (g) Asp, Tyr at position 237; (h) Glu, Ile, Met, Gin, Tyr at position 238; (i) Ile, Leu, Asn, Pro, Val at position 239; (j) Ile at position 264; (k) Phe at position 266; (m) Ala, His, Leu at position 267; (m) Asp, Glu at position 268; (o) Asp, Glu, Gly at position 271; (p) Leu at position 295; (p) Leu at position 298; (c) Glu, Phe, Ile, Leu at position 325; (r) Thr at position 326; (y) Ile, Asn at position 327; (f) Thr at position 328; (x) Lys, Arg at position 330; (c) Glu at position 331; (h) Asp at position 332; (w) Asp, Ile, Met, Val, Tyr at position 334 and (n) Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val , Trp, Tyr at position 396; according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In a further aspect of the invention, an amino acid change according to the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity is selected at each of positions selected from the group consisting of: (a) Gly, Thr in position 231; (b) Asp at position 232; (c) Trp at position 235; (d) Asn, Thr at position 236; (e) Val at position 239; (e) Asp, Glu at position 268; (g) Leu at position 295; (h) Leu at position 298; (i) Thr at position 326; (j) Lys, Arg at position 330 and (k) Lys, Met at position 396; according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asn at position 236, Glu at position 268, Lys at position 330, and Met at position 396 according to the EU numbering. In another aspect of the invention, the amino acid changes in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asn at position 236, Asp at position 268 and Lys at position 330 according to EU- numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asn at position 236, Asp at position 268, Leu at position 295, and Lys at position 330 according to the EU numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Thr at position 236, Asp at position 268 and Lys at position 330 according to EU- numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asn at position 236, Asp at position 268, Leu at position 295, Thr at position 326 and Lys at position 330 according to EU numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity are: Trp at position 235, Asn at position 236, Asp at position 268, Leu at position 295, Thr at position 326 and Lys at position 330 according to EU numbering.

In one aspect of the invention, at least one amino acid is altered according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity at at least one position selected from the group consisting of: 234 , 238, 250, 264, 267, 307 and 330, according to the EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In another aspect of the invention, one amino acid is changed in accordance with the above described method for producing a polypeptide containing a variant Fc region with increased Fc-gamma RIIb-binding activity, at position 238.

In another aspect of the invention, at least two amino acids are changed in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity, where the changes are: (i) one amino acid change at position 238 and ( II) at least one amino acid change at least one position selected from the group consisting of: 234, 250, 264, 267, 307 and 330, according to the EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In another aspect of the invention, the amino acids are changed in accordance with the method described above for obtaining a polypeptide containing an Fc region variant with increased Fc-gamma RIIb-binding activity, at one of the following positions (1)-(9): (1) at positions 234, 238, 250, 307 and 330; (2) in regulations 234, 238, 250, 264, 307 and 330; (3) in regulations 234, 238, 250, 264, 267, 307 and 330; (4) in regulations 234, 238, 250, 267, 307 and 330; (5) in regulations 238, 250, 264, 307 and 330; (6) in regulations 238, 250, 264, 267, 307 and 330; (7) in regulations 238, 250, 267, 307 and 330; (8) in regulations 238, 250 and 307; and (9) in provisions 238, 250, 307 and 330 according to the EU numbering.

In a further aspect of the invention, an amino acid change in accordance with the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity is selected at each of the positions selected from the group consisting of: (a) Tyr at position 234 ; (b) Asp at position 238; (c) Val at position 250; (d) Ile at position 264; (e) Ala at position 267; (e) Pro at position 307; and (g) Lys at position 330; according to EU numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asp at position 238 according to EU numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity are: Asp at position 238, Val at position 250 and Pro at position 307 according to EU- numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asp at position 238, Val at position 250, Pro at position 307, and Lys at position 330 according to the EU numbering. according to EU numbering. In a further aspect of the invention, the amino acid changes in accordance with the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity are: Asp at position 238, Val at position 250, Ile at position 264, Pro at position 307 and Lys at position 330 according to EU numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asp at position 238, Val at position 250, Ala at position 267, Pro at position 307 and Lys at position 330 according to EU numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity are: Tyr at position 234, Asp at position 238, Val at position 250, Pro at position 307 and Lys at position 330 according to the EU numbering. In a further aspect of the invention, the amino acid changes according to the method described above for producing a polypeptide containing an Fc region variant with enhanced Fc-gamma RIIb binding activity are: Tyr at position 234, Asp at position 238, Val at position 250, Ala at position 267, Pro at position 307 and Lys at position 330 according to EU numbering. In a further aspect of the invention, the amino acid changes in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Asp at position 238, Val at position 250, Ile at position 264, Ala at position 267, Pro at position 307 and Lys at position 330 according to EU numbering. In a further aspect of the invention, the amino acid changes in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Tyr at position 234, Asp at position 238, Val at position 250, Ile at position 264, Pro at position 307 and Lys at position 330 according to EU numbering. In a further aspect of the invention, the amino acid changes in accordance with the method described above for producing a polypeptide containing an Fc region variant with increased Fc-gamma RIIb binding activity are: Tyr at position 234, Asp at position 238, Val at position 250, Ile at position 264, Ala at position 267, Pro at position 307 and Lys at position 330 according to EU numbering. In some embodiments, the RIIb Fc-gamma has the cynomolgus Fc-gamma RIIb sequence (SEQ ID NO: 223). In some embodiments, the RIIb Fc-gamma has the sequence of a human RIIb Fc-gamma (eg, SEQ ID NO: 212, 213, or 214).

In addition, the present invention provides a method for producing a polypeptide containing a variant Fc region with increased pI compared to a polypeptide containing a parental Fc region, which consists in introducing at least two amino acid changes into the parent Fc region.

In some embodiments of the invention, a method for obtaining a polypeptide containing a variant Fc region presented in the present description, includes the implementation of the following stages, which: (a) receive a polypeptide containing the parental Fc region; (b) introducing at least two amino acid changes into the parent Fc region in the polypeptide; (c) measure the pI of the polypeptide that contains the parent Fc region and the polypeptide that contains the Fc region obtained in step (b); and (d) selecting a polypeptide containing an Fc region variant with an increased pI compared to a polypeptide containing the parental Fc region.

In one aspect of the invention, at least two amino acids are modified in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, at least two positions, selected from the group consisting of: 285, 311, 312, 315 , 318, 333, 335, 337, 341, 342, 343, 384, 385, 388, 390, 399, 400, 401, 402, 413, 420, 422 and 431, according to EU numbering. In another aspect of the invention, the resulting polypeptide contains a variant of the Fc region with increased pI, containing changes in at least two amino acid positions selected from the group consisting of: 311, 341, 343, 384, 399, 400, 401, 402 and 413, according to EU numbering.

In another aspect of the invention, at least two amino acids are modified in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, in one of the following positions (1)-(10): (1) at positions 311 and 341 ; (2) in regulations 311 and 343; (3) in regulations 311, 343 and 413; (4) in regulations 311, 384 and 413; (5) in regulations 311 and 399; (6) in regulations 311 and 401; (7) in regulations 311 and 413; (8) in regulations 400 and 413; (9) in provisions 401 and 413 and (10) in provisions 402 and 413 according to the EU numbering.

In another aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, are: Arg at position 400 and Lys at position 413 according to EU numbering. In another aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, are: Arg at position 311 and Lys at position 413 according to EU numbering. In another aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, are: Arg at position 311 and Arg at position 399 according to the EU numbering. In another aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, are: Arg at position 311 and Arg at position 343 according to the EU numbering. In another aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, are: Arg at position 311 and Arg at position 413 according to EU numbering. In another aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, are: Arg at position 311, Arg at position 343 and Arg at position 413 according to EU numbering. In another aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI, are: Arg at position 311, Arg at position 384 and Arg at position 413 according to EU numbering.

In a further aspect of the invention, amino acid changes in accordance with the method described above to obtain a polypeptide containing a variant Fc region with increased pI is selected from Lys or Arg at each position.

In a further aspect of the invention, the amino acid changes according to the preparation methods described above are selected from any of: a single change, a combination of single changes, or a combination of changes described in Tables 14-30.

Optionally, the following additional steps may be included in the methods described above to obtain a polypeptide comprising a variant Fc region, when the polypeptide further comprises an antigen-binding domain: (e) assessing the plasma pharmacokinetics of the antigen following administration of the polypeptide comprising the parental Fc region and the polypeptide containing the variant Fc regions in animals such as mice, rats, rabbits, dogs, monkeys and humans; and (e) selecting a polypeptide containing a variant Fc region with an increased ability to eliminate antigen from plasma, compared to a polypeptide containing the parental Fc region.

Polypeptides containing a variant Fc region, obtained using any of the above methods or other methods known in this field, are included in the present invention.

B. Analyzes

The anti-myostatin antibodies provided herein can be identified, screened for, or characterized for their physical/chemical properties and/or biological activities using a variety of assays known in the art.

The Fc region variants provided herein can be identified, screened for, or characterized for their physical/chemical properties and/or biological activities using a variety of assays known in the art.

1. Binding assays and other assays

In one aspect of the invention, an antibody of the invention is tested for antigen-binding activity using known methods such as ELISA, Western blotting, BIACORE®, etc. In one aspect of the invention, a polypeptide containing a variant Fc region of the invention is tested for binding activity to its Fc receptor using known methods such as BIACORE®, etc.

According to another aspect of the invention, competition assays can be used to identify an antibody that competes for binding to myostatin with any of the anti-myostatin antibodies described herein. In some embodiments of the invention, when the specified competitive antibody is present in excess, it blocks (for example, reduces) the binding of the reference antibody to myostatin by at least 10%, 15%, 20%, 25%, 30%, 35%, 40 %, 45%, 50%, 55%, 60%, 65%, 70%, 75% or more. In some cases, binding is inhibited by at least 80%, 85%, 90%, 95% or more. In some embodiments, said competing antibody binds to the same epitope (e.g., a linear or conformational epitope) that the anti-myostatin antibody provided herein binds (e.g., the anti-myostatin antibody described in Tables 2a, 11a, or 13). In other aspects of the invention, the reference antibody has a VH and VL pair as described in Table 2a, 11a, or 13. For a detailed description of exemplary methods for mapping the epitope to which the antibody binds, see Morris, Epitope Mapping Protocols, in: Methods in Molecular Biology, Vol. 66, Humana Press, Totowa, NJ, 1996.

As an example of a competitive assay, an immobilized latent myostatin or myostatin propeptide is incubated in a solution containing a first labeled antibody that binds to myostatin and a second unlabeled antibody to be tested for its ability to compete with the first antibody for binding to myostatin. The second antibody may be present in the hybridoma supernatant. As a control, immobilized myostatin is incubated in a solution containing the first labeled antibody but not the second unlabeled antibody. After incubation under conditions suitable for binding of the first antibody to myostatin, the excess of unbound antibody is removed and the amount of label associated with the immobilized myostatin is measured. If the amount of label associated with immobilized myostatin is significantly reduced in the test sample compared to the control sample, then this indicates that the second antibody competes with the first antibody for binding to myostatin (see, for example, Harlow and Lane, Antibodies: A Laboratory Manual, Chapter 14, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1988).

Assays for determining the binding activity of a polypeptide containing a variant Fc region to one or more Fc-gamma R members are provided herein or are known in the art. Such binding assays include, but are not limited to, the BIACORE® assay using the Surface Plasmon Resonance (SPR) phenomenon, Amplified Luminescent Proximity Homogeneous Assay (ALPHA), ELISA , and cell sorting by fluorescence intensity (FACS) (Lazar et al., Proc. Natl. Acad. Sci. USA 103 (11), 2006, pp. 4005-4010).

In one embodiment, the BIACORE® assay can be used to determine whether the binding activity of a polypeptide containing a variant Fc region is increased, or is maintained at the same level, or is reduced against a particular member of the Fc-gamma R family. For example, for this purpose, it is determined whether the value of dissociation constant (KD) decreases or increases, which is determined using sensorgram analysis when various Fc-gamma R are reacted as an analyte with polypeptides containing a variant Fc region, immobilized or "captured" on a sensor chip, using known methods and reagents, such as protein A, protein L, protein A/G, protein G, antibodies to the lambda chain, antibodies to the kappa chain, antigenic peptides, antigenic proteins. Changes in binding activity can also be determined by comparing changes in the number of resonance units (RU) in the sensorgram before and after one or more types of Fc-gamma R are reacted as analytes with "captured" polypeptides containing a variant Fc region. Alternatively, the Fc-gamma R can be immobilized or "captured" on sensor chips and polypeptides containing the variant Fc region used as the analyte.

When performing BIACORE® assays, one of the substances (ligand) intended to study the interaction is immobilized on a thin layer of gold on a sensor chip and illuminated by light from the rear surface of the sensor chip in such a way that there is a complete reflection at the interface between the thin layer of gold and glass, a region with reduced intensity (SPR signal) is formed in part of the reflected light. Prepare another substance (analyte) intended to study the interaction, for injection on the surface of the sensor chip; and when the ligand binds to the analyte, the mass of the immobilized ligand molecule increases and the refractive index of the solvent on the surface of the sensor chip changes. As a result of said refractive index change, the position of the SPR signal shifts (and vice versa, the position of the signal returns to its original position if dissociation of said binding occurs). With the BIACORE® system, the level of the above-described shift, or more specifically the change in mass versus time, is determined by plotting the change in mass on the surface of the sensor chip along the vertical axis, and thus obtaining quantitative data (sensogram). The amount of analyte bound to the ligand immobilized on the surface of the sensor chip is determined from the sensorgram. Kinetic parameters such as association rate constant (ka) and dissociation rate constant (kd) are determined from the sensory curves, and affinity (KD) is determined as the ratio of these constants. As a method for inhibition assay, the BIACORE® method is preferably used. Examples of such a method for inhibition assays are described in Lazar et al., Proc. Natl. Acad. sci. USA 103(11), 2006, pp. 4005-4010.

ALPHA-based screening is based on ALPHA technology, which is based on the principle described below, using two types of beads, donors and acceptors. Luminescent signals are detectable only when the molecules associated with the donor beads physically interact with the molecules associated with the acceptor beads and when both beads are in close proximity to each other. Excited by a laser beam, the photosensitizer in donor granules converts environmental oxygen into excited singlet oxygen. When singlet oxygen diffuses from the donor granules and reaches the acceptor granules located in close proximity, a chemiluminescent reaction is induced in the granules, which ultimately leads to the emission of light. If the molecules bound to the donor beads do not interact with the molecules bound to the acceptor beads, then the chemiluminescent reaction does not occur because the singlet oxygen that is produced by the donor beads does not reach the acceptor beads.

For example, a biotinylated polypeptide complex is coupled to donor beads and a glutathione-8-transferase (GST) labeled Fc-gamma receptor is coupled to acceptor beads. In the absence of a competing polypeptide complex containing a variant Fc region, the polypeptide complex containing the parent Fc region interacts with the Fc gamma receptor and generates signals at a wavelength of 520 to 620 nm. The polypeptide complex containing the unlabeled Fc region variant competes with the polypeptide complex containing the parent Fc region for binding to the Fc gamma receptor. Relative binding affinity can be assessed by quantifying the decrease in fluorescence due to competition. Methods for biotinylating polypeptide complexes such as antibodies with sulfo-NHS-biotin or the like are well known. Method for Expressing an Fc-gamma Receptor and a GST in a Cell Carrying a Fusion Gene Obtained by Fusing a Polynucleotide That Encodes an Fc-gamma Receptor in Frame with a Polynucleotide That Encodes GST in an Expression Vector and Performing Purification with a Glutathione-Containing Column , are adapted accordingly, obtaining a method of labeling the Fc-gamma receptor with GST. The induced signals can be analyzed, for example, by fitting to a single site competition model based on non-linear regression analysis using software such as GRAPHPAD PRISM (GraphPad; San Diego).

The term "variant Fc region with reduced Fc-gamma R-binding activity" refers to an Fc region that binds to an Fc-gamma R with significantly lower binding activity than the parent Fc region when assays are performed using substantially the same amounts corresponding parent Fc region and variant Fc region. In addition, the term "variant Fc region with enhanced Fc gamma R binding activity" refers to an Fc region that binds to an Fc gamma R with substantially stronger binding activity than the parent Fc region when assayed using substantially equal amounts of the corresponding parental Fc region and the variant Fc region. The term "variant Fc region with retained Fc-gamma R binding activity" refers to an Fc region that binds to an Fc-gamma R with activity equivalent to or substantially the same as that of the parent Fc region when assayed with using substantially equal amounts of the corresponding parental Fc region and the polypeptide containing the variant Fc region.

Whether the binding activity of the Fc region to various Fc gamma Rs increases or decreases can be determined by an increase or decrease in the level of binding of various Fc gamma Rs to the Fc region when determined using the measurement method described above. In the context of the present description, the level of binding of various Fc-gamma Rs to the Fc region can be estimated as a value obtained by dividing the difference in RU levels in the sensorograms before and after the interaction of various Fc-gamma Rs, which are used as an analyte, with the Fc region, on the difference in RU levels on the sensorgrams, which changed before and after the “capture” of the Fc regions on the sensor chips.

According to the present invention, increased Fc-gamma RIIb binding activity preferably means, for example, that the ratio [KD value of the parent Fc region for Fc-gamma RIIb]/[KD value of the variant Fc region for Fc-gamma RIIb] when measured as values The KD measured using the above method is preferably 2.0 or more, 3.0 or more, 4.0 or more, 5.0 or more, 6.0 or more, 7.0 or more, 8.0 or more, 9.0 or more, 10 or more, 15 or more, 20 or more, 25 or more, 30 or more, 35 or more, 40 or more, 45 or more, or even 50 or more, 55 or more, 60 or more more, 65 or more, 70 or more, 75 or more, 80 or more, 85 or more, 90 or more, 95 or more, or 100 or more.

The KD value (mol/l) of the variant Fc region specified in the present description for Fc-gamma RIIb is preferably lower than the parent Fc region for Fc-gamma RIIb, and may be, for example, 2.0×10 ^-6 M or less, 1.0×10 ^-6 M or less, 9.0×10 ^-7 M or less, 8.0×10 ^-7 M or less, 7.0×10 ^-7 M or less, 6.0 x10 ^-7 M or less, 5.0 x 10 ^-7 M or less, 4.0 x 10 ^-7 M or less, 3.0 x 10 ^-7 M or less, 2.0 x 10 ^-7 M or less, 1.0×10 ^-7 M or less, 9.0×10 ^-8 M or less, 8.0×10 ^-8 M or less, 7.0×10 ^-8 M or less, 6.0× 10 ^-8 M or less, 5.0×10 ^-8 M or less.

In addition, a variant Fc region with increased binding selectivity to Fc-gamma RIIb compared to Fc-gamma RIIa means an Fc region in which: (a) Fc-gamma RIIb-binding activity is increased and Fc-gamma RIIa-binding activity remains at this level or decreases, (b) Fc-gamma RIIb-binding activity increases and Fc-gamma RIIa-binding activity also increases, but the degree of increase in Fc-gamma RIIa-binding activity is lower than the degree of increase in Fc-gamma RIIb- binding activity; or (c) Fc-gamma RIIb binding activity is reduced and Fc-gamma RIIa binding activity is also reduced, but the reduction rate of Fc-gamma RIIb binding activity is lower than the reduction rate of Fc-gamma RIIa binding activity. Whether the Fc region variant has increased binding selectivity for the Fc gamma RIIb relative to the Fc gamma RIIa can be determined, for example, by comparing the ratio of the KD value for the Fc gamma RIIa to the KD value for the Fc gamma RIIb variant Fc region (ratio [K.D value of Fc-region variant for Fc-gamma RIIa]/[KD value of Fc-region variant for Fc-gamma RIIb]), with the ratio of KD value for Fc-gamma RIIa and KD value for Fc-gamma RIIb parent Fc -region (the ratio of [KD value of the parent Fc region for Fc-gamma RIIa]/[KD value of the parent Fc region for Fc-gamma RIIb]), which is determined according to the above examples. In particular, when the KD ratio of the Fc region variant is higher than that of the parent Fc region, then the Fc region variant can be considered to have increased binding selectivity for Fc gamma RIIb relative to Fc gamma RIIa relative to the parent Fc region. . With respect to human receptors, Fc-gamma RIIb binding activity is likely to correlate with Fc-gamma RIIa (R-type) binding activity rather than Fc-gamma RIIa (H-type), since the amino acid sequence of Fc-gamma RIIb is more identity with Fc-gamma RIIa (R-type) than with Fc-gamma RIIa (H-type). Thus, the identified amino acid change(s) that can(may) increase binding selectivity to human Fc-gamma RIIb compared to human Fc-gamma RIIa (R-type) is(are) ) important(s) for increasing the selectivity of binding to Fc-gamma RIIb compared to Fc-gamma RIIa in humans.

When a variant Fc region of the present invention has increased binding activity to Fc-gamma RIIb and reduced binding activity to Fc-gamma RIII (such as Fc-gamma RIIIa or Fc-gamma RIIIb) compared to the parental Fc-region, then the Fc region variant can be considered to be Fc-gamma RIIb-specific.

2. Activity assays

One of the objects of the invention are assays designed to identify antibodies to myostatin, which have biological activity. The biological activity may include, for example, inhibition of myostatin activation, blockade of the release of mature myostatin from latent myostatin, inhibition of proteolytic cleavage of latent myostatin, blockade of the availability of latent myostatin to protease, and the like. Also provided are antibodies having said biological activity in vivo and/or in vitro. In some embodiments, an antibody of the invention is tested for a specified biological activity.

In some embodiments, whether a test antibody has the ability to inhibit latent myostatin cleavage is determined by detecting the latent myostatin cleavage product (myostatin) using a method known in the art such as electrophoresis, chromatography, immunoblot analysis, enzyme-linked immunosorbent assay (ELISA) or mass spectrometry, after contacting a protease that can cleave latent myostatin with latent myostatin with or without the test antibody (see, for example, Thies et al., Growth Factors 18 (4), 2001, pp. 251-259 ( 2001)). When a reduced amount of latent myostatin cleavage product (eg, human myostatin propeptide) is detected in the presence of (or after contact with) the test antibody, the test antibody is identified as an antibody that can inhibit the breakdown of latent myostatin. In some embodiments, whether a test antibody has the ability to block access to latent myostatin is determined using methods to evaluate protein interactions between protease and latent myostatin, for example, using ELISA or BIACORE®. When a decrease in the interaction between the protease and latent myostatin is detected in the presence of (or after contact with) the test antibody, the test antibody is identified as an antibody that can block the access of the protease to latent myostatin.

In some embodiments, whether a test antibody has the ability to block the release of mature myostatin from latent myostatin is determined by determining the activity of mature myostatin, e.g., myostatin receptor binding activity, or the ability to mediate signal transduction in a cell expressing the myostatin receptor (e.g., ActRIIb ). Cells that can be used in such an assay may be cells that express an endogenous receptor for myostatin, such as L6 myocytes, or may be genetically modified, transiently or stably, such that they express a transgene encoding a receptor for myostatin. , a receptor for activin, such as the type II activin receptor (Thies et al., supra). The binding of myostatin to the myostatin receptor can be assessed using a receptor binding assay. Myostatin-mediated signal transduction can be determined at any level in the signal transduction pathway, for example, by measuring Smad polypeptide phosphorylation, measuring the expression of a myostatin-regulated gene, including a reporter gene, or measuring the proliferation of myostatin-dependent cells. By reducing the activity of mature myostatin in the presence of the test antibody (or after contact with it), the test antibody is identified as an antibody that can block the release of mature myostatin from latent myostatin.

Inhibition of myostatin activation can also be detected and/or measured using the methods set forth as an example in the described embodiments of the invention. Using these assays, or other suitable assay types, test antibodies can be screened for their ability to inhibit myostatin activation. In some embodiments, the inhibition of myostatin activation comprises a reduction in myostatin activation of at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40% or more in this assay compared to a negative control in the same conditions. In some embodiments, it includes inhibiting myostatin activation by at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% or more.

Another object of the invention are assays designed to identify antibodies to myostatin, which form an immune complex (ie, an antigen-antibody complex) with myostatin. Also provided are antibodies having said biological activity in vivo and/or in vitro.

In some embodiments, an antibody of the invention is tested for a specified biological activity.

In some embodiments of the invention, the formation of the immune complex is assessed using a method such as gel filtration chromatography, ultracentrifugation, light scattering, electron microscopy or mass spectrometry (Mol. Immunol. 39, 2002, cc. 77-84, Mol. Immunol. 47, 2009, pp. 357-364). The possibility of implementing these methods is based on the fact that the immune complex is a larger molecule than an individual antibody or individual antigen. If a large complex containing two or more antibodies and two or more antigens (e.g., myostatin molecules) is detected in the presence of a test antibody and an antigen, then the test antibody is identified as an antibody that can form an immune complex containing two or more antibodies. and two or more myostatin molecules. In another embodiment of the invention, immune complex formation is assessed using a method such as ELISA, FACS, or SPR (Surface Plasmon Resonance Analysis; e.g., BIACORE™) (Shields et al., J. Biol. Chem. 276(9) , 2001, pp. 6591-6604; Singh et al., J. Immunol. Methods 50, 1982, pp. 109-114; Suzuki et al., J. Immunol. 184(4), 2010, pp. 1968-1976 ; Luo et al., mAbs 1 (5), 2009, pp. 491-504). The possibility of using these methods is based on the fact that an immune complex containing two or more antibodies and two or more antigens can bind more strongly to the Fc receptor or a component of the complement system than an antibody or antigen when used alone. If increased binding to the Fc receptor or a component of the complement system is found in the presence of both the test antibody and the antigen compared to the presence of the antibody alone, then the test antibody is identified as an antibody that can form an immune complex containing two or more antibodies and two or more number of myostatin molecules. In another embodiment of the invention, immune complex formation is assessed by administering a test antibody to an animal (eg a mouse) and measuring the clearance of the antigen from plasma. As described above, an antibody that forms an immune complex containing two or more antibodies with two or more antigens is expected to accelerate the elimination of antigens from plasma. Thus, if accelerated clearance of myostatin from plasma is found in mice injected with a test antibody compared to mice injected with a reference antibody, then the test antibody is identified as an antibody that can form an immune complex containing two or more antibodies and two or more myostatin molecules, more effective than the reference antibody.

D. Immunoconjugates

Some embodiments of the invention are immunoconjugates comprising a myostatin antibody as described herein conjugated to one or more cytotoxic agents such as chemotherapeutic agents or drugs, growth inhibitory agents, toxins (e.g., protein toxins, enzymatically active bacterial, fungal, , of plant or animal origin or their fragments) or radioactive isotopes.

In one embodiment, the immunoconjugate is an antibody-drug conjugate (ADC) in which the antibody is conjugated to one or more drugs, including (but not limited to) maytansinoid (see US Nos. 5208020, 5416064 and EP 0425235 B1); auristatin, for example, fragments DE and DF of monomethylauristatin (MMAE and MMAF) (see US No. 5635483, 5780588 and 7498298); dolastatin; calicheamicin or derivatives thereof (see US Nos. 5712374, 5714586, 5739116, 5767285, 5770701, 5770710, 5773001 and 5877296; Hinman et al., Cancer Res. 53, 1993, pp. 3336-3342; and Lode et al. Cancer Res. 58, 1998, pp. 2925-2928); an anthracycline such as daunomycin or doxorubicin (see Kratz et al., Curr. Med. Chem. 13, 2006, pp. 477-523; Jeffrey et al., Bioorg. Med. Chem. Lett. 16, 2006, pp. 358-362; Torgov et al., Bioconjug. Chem. 16, 2005, pp. 717-721; Nagy et al., Proc. Natl. Acad. Sci. USA 97, 2000, pp. 829-834; Dubowchik et al. ., Bioorg. & Med. Chem. Letters 12, 2002, pp. 1529-1532; King et al., J. Med. Chem. 45, 2002, pp. 4336-4343; and US No. 6630579); methotrexate; vindesine; a taxane such as docetaxel, paclitaxel, larotaxel, tesetaxel and orthotaxel; trichothecene and CC1065.

In another embodiment, the immunoconjugate comprises an antibody as described herein conjugated to an enzymatically active toxin or fragment thereof, including (but not limited to) diphtheria toxin A chain, non-binding diphtheria toxin active moieties, exotoxin A chain ( from Pseudomonas aeruginosa), ricin A-chain, abrin A-chain, modecin A-chain, alpha-sarcin, Aleurites fordii proteins, diantine proteins, Phytolaca americana proteins (PAPI, RARP and PAP-5), inhibitor from Momordica charantia, curcin , crotin, an inhibitor from Sapaonaria officinalis, gelonin, mitogellin, restrictocin, fenomycin, enomycin, and trichothecenes.

In another embodiment, the immunoconjugate comprises an antibody as described herein conjugated to a radioactive atom to form a radioconjugate. A wide variety of radioactive isotopes can be used to prepare radioconjugates. Examples include ²¹¹ At, ¹³¹ I, ¹²⁵ I, ⁹⁰ Y, ¹⁸⁶ Re, ¹⁸⁸ Re, ¹⁵³ Sm, ²¹² Pb, ³² P, ²¹² Pb and radioactive isotopes of Lu. When the radioconjugate is used for detection, it may contain a radioactive atom for scintigraphy, such as TC ^99m or I ¹²³ , or a spin label for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI), such as iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese or iron.

Antibody-cytotoxic agent conjugates can be prepared using a wide variety of bifunctional protein-binding agents such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC ), iminothiolane (IT), difunctional imidoester derivatives (such as dimethyl adipimidate×HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bisazido derivatives (such as bis(p-azidobenzoyl)hexanediamine), derivatives bisdiazonium (such as bis(para-diazoniumbenzoyl)ethylenediamine), diisocyanates (such as toluene-2,6-diisocyanate), and dual activity fluoro compounds (such as 1,5-difluoro-2,4-dinitrobenzene). For example, the ricin immunotoxin can be prepared according to the method described in Vitetta et al., Science 238, 1987, cc. 1098-1104. C14 labeled 1-isothiocyanatebenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an example of a chelating agent used to conjugate a radionucleotide to an antibody (see WO 94/11026). The linker may be a "cleavable linker" to facilitate release of the cytotoxic drug. For example, an acid-labile linker, a peptidase-sensitive linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker can be used (Chad et al., Cancer Res. 52, 1992, pp. 127-131; US No. 5208020).

In the context of the present description, immunoconjugates or ADCs mean (but not limited to) these conjugates obtained using cross-linking reagents, which include (but not limited to) BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC and sulfo-SMPB, and SVSB (succinimidyl(4- vinylsulfone)benzoate) commercially available (for example, from Pierce Biotechnology, Inc., Rockford, Illinois, USA).

E. Methods and compositions for diagnosing and detecting

In some embodiments, any of the anti-myostatin antibodies provided herein can be used to detect the presence of myostatin in a biological sample. The concept of "detection (detection)" in the context of the present description provides for quantitative or qualitative detection. In one embodiment, the biological sample comprises a cell or tissue, e.g., serum, whole blood, plasma, biopsy sample, tissue sample, cell suspension, saliva, sputum, oral fluid, cerebrospinal fluid, amniotic fluid, ascitic fluid, milk, colostrum, mammary secretion, lymph, urine, sweat, lacrimal fluid, gastric juice, synovial fluid, peritoneal fluid, lens fluid or mucus.

One embodiment of the invention is an anti-myostatin antibody provided herein for use in a diagnostic or detection method. Another object of the invention relates to a method for detecting the presence of latent myostatin or myostatin propeptide in a biological sample. In another embodiment, the method comprises contacting a biological sample with an anti-myostatin antibody provided herein under conditions that allow the anti-myostatin antibody to bind to myostatin, and detecting the formation of a complex between the anti-myostatin antibody and myostatin. Said method may be an in vitro or in vivo method. In one embodiment, the anti-myostatin antibody is used to select individuals who can be treated with the anti-myostatin antibody, for example, where myostatin is a biomarker for patient selection.

Examples of disorders that can be diagnosed using an antibody of the invention include, but are not limited to, muscular dystrophy (MD; including Duchenne muscular dystrophy), amyotrophic lateral sclerosis (ALS), muscle atrophy, organ atrophy, carpal tunnel syndrome , frailty, congestive obstructive pulmonary disease (COPD), sarcopenia, cachexia, muscle wasting syndromes, HIV-induced muscle wasting, type 2 diabetes, impaired glucose tolerance, metabolic syndrome (including syndrome X), insulin resistance (including resistance induced trauma, e.g., burns or nitrogen imbalance), adipose tissue disorders (e.g., obesity, dyslipidemia, non-alcoholic fatty liver disease, etc.), osteoporosis, osteopenia, osteoarthritis, and metabolic bone disorders (including low bone mass , premature ovarian failure, androgen suppression, vitamin deficiency D, secondary hyperparathyroidism, nutritional deficiencies, and anorexia nervosa).

Some embodiments of the invention are labeled antibodies to myostatin. Labels include, but are not limited to, labels or moieties that can be detected directly (such as fluorescent, chromophoric, electron dense, chemiluminescent, and radioactive labels) as well as moieties such as enzymes or ligands that are indirectly detectable, such as through an enzymatic reaction or molecular interaction. Examples of labels include, but are not limited to, ³² P, ¹⁴ C, ¹²⁵ I, ³ H, and ¹³¹ I radioisotopes, fluorophores such as rare earth metal chelates or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, umbelliferone, luciferases , e.g., firefly luciferase and bacterial luciferase (US No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, horseradish peroxidase (HRP), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, saccharide oxidases, e.g., glucose oxidase, galactose oxidase, and glucose -6-phosphate dehydrogenase, heterocyclic oxidases such as uricase and xanthine oxidase linked to an enzyme that uses hydrogen peroxide to oxidize a dye precursor such as HRP, lactoperoxidase or microperoxidase, biotin/avidin, spin labels, bacteriophage labels, stable free radicals and so on.

E. Pharmaceutical compositions

Pharmaceutical Sciences, 16-е изд. под ред. Osol А., 1980) в форме лиофилизированных композиций или водных растворов.Pharmaceutical compositions of the anti-myostatin antibody provided herein are prepared by mixing said antibody, having the required degree of purity, with one or more optional pharmaceutically acceptable carriers (

Pharmaceutical Sciences, 16th ed. ed. Osol A., 1980) in the form of lyophilized compositions or aqueous solutions.

Pharmaceutical compositions of a polypeptide containing an Fc region variant as described herein are prepared by mixing said polypeptide having the desired purity with one or more optional pharmaceutically acceptable carriers in the form of lyophilized compositions or aqueous solutions.

Pharmaceutically acceptable carriers are generally non-toxic to recipients at the dosages and concentrations employed and include (but are not limited to): buffers such as phosphate, citrate, and other organic acid buffers; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol; butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol and meta-cresol); low molecular weight polypeptides (containing less than about 10 residues); proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as poly(vinylpyrrolidone); amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal-containing complexes (eg Zn-protein complexes); and/or nonionic surfactants such as polyethylene glycol (PEG). In the context of the present description, examples of pharmaceutically acceptable carriers also include interstitial drug dispersing agents such as soluble, neutrally active hyaluronidase glycoproteins (sHASEGP), for example, human soluble hyaluronidase glycoproteins PH-20, such as rHuPH20 (HYLENEX ^® , Baxter International, Inc.). Some examples of sHASEGP and methods of using them, including rHuPH20, are described in US Patent Publication Nos. 2005/0260186 and 2006/0104968. According to one aspect of the invention, sHASEGP is combined with one or more additional glycosaminoglycanases, such as chondroitinases.

Examples of lyophilized antibody compositions are described in US No. 6267958. Aqueous compositions of antibodies include those described in US No. 6171586 and WO 2006/044908, the latter compositions include histidine acetate buffer.

The composition according to the invention may also contain more than one active substance, if necessary for the specific indication to be treated, preferably substances with complementary activities that do not adversely affect each other. Such active substances must be present in combination in amounts effective to achieve the intended purpose.

Pharmaceutical Sciences, 16-е изд., под ред. Osol А., 1980.The active substances can also be included in microcapsules, for example, obtained by coacervation or interfacial polymerization methods, for example in hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively, in colloidal drug delivery systems (for example, in liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules) or in macroemulsion. Such methods are described in

Pharmaceutical Sciences, 16th ed., ed. Osol A., 1980.

Sustained release formulations can be prepared. Suitable examples of sustained release preparations are semi-permeable matrices of solid hydrophobic polymers containing an antibody or conjugate, such matrices are shaped articles such as films or microcapsules.

Compositions intended for use in vivo, as a rule, must be sterile. Sterility can easily be achieved, for example, by filtration through sterile filter membranes.

G. Therapeutic Methods and Compositions

Any of the anti-myostatin antibodies provided herein can be used in therapeutic methods. Likewise, any of the polypeptides containing the Fc region variant provided herein can be used in therapeutic methods.

One object of the invention is an antibody to myostatin, presented in the present description, for use as a medicine. Another object of the invention is an antibody to myostatin, presented in the present description, for use in the treatment of muscle wasting syndrome. In some embodiments, an anti-myostatin antibody for use in a method of treatment is provided. In some embodiments, an anti-myostatin antibody is provided for use in a method of treating a subject having muscle wasting syndrome, comprising administering to the subject an effective amount of the anti-myostatin antibody. In one of these embodiments of the invention, the method also includes the introduction to the individual in an effective amount of at least one additional therapeutic agent. In other embodiments, the anti-myostatin antibody is for use in increasing muscle mass. In some embodiments, the anti-myostatin antibody is for use in a method for increasing muscle mass in a subject, comprising administering to the subject an effective amount of the anti-myostatin antibody to increase muscle mass. In further embodiments, the anti-myostatin antibody is for use in increasing muscle tissue strength. In some embodiments, the anti-myostatin antibody is for use in a method of increasing muscle strength in a subject, comprising administering to the subject an effective amount of the anti-myostatin antibody to increase muscle strength. In further embodiments, the anti-myostatin antibody is for use in reducing body fat accumulation. In some embodiments, the anti-myostatin antibody is for use in a method for reducing body fat accumulation in a subject, comprising administering to the subject an effective amount of an anti-myostatin antibody to reduce body fat accumulation. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

An anti-myostatin antibody of the present invention may have pH dependent binding characteristics. In other embodiments, the invention is an anti-myostatin antibody for use in increasing the clearance of myostatin from plasma. In some embodiments, an anti-myostatin antibody is provided for use in a method of increasing plasma clearance of myostatin in a subject, comprising administering to the subject an effective amount of an anti-myostatin antibody to increase plasma clearance of myostatin. In one embodiment, an anti-myostatin antibody with pH-dependent binding characteristics increases the clearance of myostatin from plasma compared to a canonical anti-myostatin antibody that does not have pH-dependent binding characteristics. In a further embodiment of the invention, an anti-myostatin antibody with pH-dependent binding characteristics, having the ability to bind at a pH between pH 5.8 and pH 7.4, increases the clearance of myostatin from plasma compared to a canonical anti-myostatin antibody that does not have a pH- dependent binding characteristics. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

The next object of the invention is the use of antibodies to myostatin for the preparation or production of drugs. In one of the embodiments of the invention, the drug is for the treatment of muscle wasting syndrome. In another embodiment, the drug is used in a method for treating muscle wasting syndrome, comprising administering to an individual having muscle wasting syndrome an effective amount of the drug. In one of these embodiments of the invention, the method also includes the introduction to the individual in an effective amount of at least one additional therapeutic agent. In a further embodiment of the invention, the drug is intended to increase muscle tissue mass. In another embodiment, the drug is for use in a method for increasing muscle mass in an individual, comprising administering to the individual an effective amount of the drug to increase muscle mass. In a further embodiment of the invention, the drug is intended to increase the strength of muscle tissue. In another embodiment, the drug is for use in a method for increasing muscle strength in a subject, comprising administering to the subject an effective amount of a drug to increase muscle strength. In a further embodiment of the invention, the drug is for reducing the accumulation of body fat. In another embodiment, the drug is for use in a method for reducing body fat accumulation in an individual, comprising administering to the individual an effective amount of a drug to reduce body fat accumulation. An "individual" in the context of any of the above embodiments of the invention may be a human.

An anti-myostatin antibody of the present invention may have pH dependent binding characteristics. In a further embodiment of the invention, the drug is intended to increase the clearance of myostatin from plasma. In another embodiment, the drug is for use in a method of increasing plasma myostatin clearance in a subject, comprising administering to the subject an effective amount of the drug to increase plasma myostatin clearance. In one embodiment, an anti-myostatin antibody with pH-dependent binding characteristics increases the clearance of myostatin from plasma compared to a canonical anti-myostatin antibody that does not have pH-dependent binding characteristics. In another embodiment, the anti-myostatin antibody has different pH dependent binding characteristics between pH 5.8 and pH 7.4. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

Another object of the invention is a method for the treatment of muscle wasting syndrome. In one embodiment, the method comprises administering to an individual having said muscle wasting syndrome an effective amount of an anti-myostatin antibody provided herein. In one of the embodiments of the invention, the method also includes the introduction to the individual in an effective amount of at least one additional therapeutic agent. An "individual" in the context of any of the above embodiments of the invention may be a human.

Another object of the invention is a method for increasing the mass of muscle tissue in an individual. In one embodiment, the method comprises administering to an individual an effective amount of an anti-myostatin antibody provided herein to increase muscle mass. In one embodiment, the "individual" is a human.

Another object of the invention is a method for increasing the strength of muscle tissue in an individual. In one embodiment, the method comprises administering to an individual an effective amount of an anti-myostatin antibody provided herein to increase muscle tissue strength. In one embodiment, the "individual" is a human.

Another object of the invention is a method for reducing the accumulation of body fat in an individual. In one embodiment, the method comprises administering to an individual an effective amount of an anti-myostatin antibody provided herein to reduce accumulation of body fat. In one embodiment, the "individual" is a human.

An anti-myostatin antibody of the present invention may have pH dependent binding characteristics. Another embodiment of the invention is a method for increasing the plasma clearance of myostatin in an individual. In one embodiment, the method comprises administering to the subject an effective amount of an anti-myostatin antibody provided herein to increase clearance of myostatin from plasma. In one embodiment, an anti-myostatin antibody with pH-dependent binding characteristics increases the clearance of myostatin from plasma compared to a canonical anti-myostatin antibody that does not have pH-dependent binding characteristics. In another embodiment, the anti-myostatin antibody has different pH dependent binding characteristics between pH 5.8 and pH 7.4. In one embodiment, the "individual" is a human.

Another object of the invention are pharmaceutical compositions containing any of the antibodies to myostatin presented in the present description, which are intended, for example, for use in any of the above therapeutic methods. In one embodiment, the pharmaceutical composition comprises any of the anti-myostatin antibodies provided herein and a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition comprises any of the anti-myostatin antibodies provided herein and at least one additional therapeutic agent.

The next object of the invention is a pharmaceutical composition intended for the treatment of muscle wasting syndrome. In another embodiment of the invention, the pharmaceutical composition is intended to increase the mass of muscle tissue. In another embodiment of the invention, the pharmaceutical composition is intended to increase the strength of muscle tissue. In a further embodiment of the invention, the pharmaceutical composition is for reducing the accumulation of body fat. An anti-myostatin antibody of the present invention may have pH dependent binding characteristics. In a further embodiment of the invention, the pharmaceutical composition is for increasing the clearance of myostatin from plasma. In one of the embodiments of the invention, the pharmaceutical composition is administered to an individual who has a muscle wasting syndrome. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

A further aspect of the invention are methods of preparing a medicament or pharmaceutical composition comprising admixing any of the anti-myostatin antibodies provided herein with a pharmaceutically acceptable carrier, for example, for use in any of the above therapeutic methods. In one of the embodiments of the invention, methods for preparing a drug or pharmaceutical composition further include adding at least one additional therapeutic agent to the drug or pharmaceutical composition.

In some embodiments, muscle wasting is selected from the group including muscular dystrophy (MD; including Duchenne muscular dystrophy), amyotrophic lateral sclerosis (ALS), muscle atrophy, organ atrophy, carpal tunnel syndrome, frailty, congestive obstructive pulmonary disease (COPD), sarcopenia, cachexia, muscle wasting syndromes, HIV-induced muscle wasting, type 2 diabetes, impaired glucose tolerance, metabolic syndrome (including syndrome X), insulin resistance (including resistance induced by trauma, such as burn or nitrogen imbalance) , adipose tissue disorders (eg, obesity, dyslipidaemia, non-alcoholic fatty liver disease, etc.), osteoporosis, osteopenia, osteoarthritis, and metabolic bone disorders (including low bone mass, premature ovarian failure, androgen suppression, vitamin D deficiency, secondary hyperparathyroidism, nutritional deficiencies and vomit anorexia).

Any of the polypeptides containing the Fc region variant provided herein can be used in therapeutic methods. Another object of the invention are pharmaceutical compositions containing a polypeptide that contains any of the polypeptides containing variants of the Fc regions presented in the present description, for example, for use in therapeutic methods. In one of the embodiments of the invention, the pharmaceutical composition includes a polypeptide that contains any of the polypeptides containing variants of the Fc regions presented in the present description, and a pharmaceutically acceptable carrier. In another embodiment of the invention, the pharmaceutical composition contains a polypeptide that contains any of the variants of the Fc regions presented in the present description, and at least one additional therapeutic agent.

One of the objects of the invention is a polypeptide containing a variant Fc region, intended for use as a drug. Another aspect of the invention is a polypeptide containing a variant Fc region for the treatment of a disorder. In some embodiments, the invention is a polypeptide containing a variant Fc region for use in a method of treatment. In some embodiments, the invention is a polypeptide comprising an Fc region variant for use in a method of treating an individual who has a disorder, comprising administering to the individual an effective amount of a polypeptide comprising the Fc region variant provided herein. In one of the embodiments of the invention, the method further includes the introduction to the individual in an effective amount of at least one additional therapeutic agent. In one embodiment, the "individual" is a human.

Another object of the invention is the use of a polypeptide containing a variant Fc region for the preparation or production of a drug. In one embodiment, the drug is for the treatment of a disorder. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the drug is for use in a method for treating a disorder, comprising administering to an individual who has the disorder being treated, an effective amount of the drug. In one of the embodiments of the invention, the method further includes the introduction to the individual in an effective amount of at least one additional therapeutic agent. In one embodiment, the "individual" is a human.

The next object of the invention is a method for treating disorders. In one embodiment, the method comprises administering to an individual who has said disorder an effective amount of a polypeptide comprising a variant Fc region. In one of the embodiments of the invention, the method further includes the introduction to the individual in an effective amount of at least one additional therapeutic agent. In one embodiment, the "individual" is a human.

A further aspect of the invention are pharmaceutical compositions that comprise a polypeptide containing a variant Fc region provided herein for use in a therapeutic method, such as any of the therapeutic methods provided herein. In one of the embodiments of the invention, the pharmaceutical composition contains a polypeptide that contains a variant of the Fc region presented in the present description, and a pharmaceutically acceptable carrier. In another embodiment of the invention, the pharmaceutical composition contains a polypeptide that contains a variant Fc region, presented in the present description, and at least one additional therapeutic agent.

According to a further aspect of the invention, the pharmaceutical composition is for the treatment of a disorder. In one embodiment of the invention, the pharmaceutical composition is administered to an individual who has a disorder. In one embodiment, the "individual" is a human.

In other embodiments, a polypeptide comprising a variant Fc region is for use in suppressing the activation of B cells, mast cells, dendritic cells, and/or basophils. In some embodiments, the polypeptide comprising the Fc region variant is for use in a method for suppressing the activation of B cells, mast cells, dendritic cells, and/or basophils in an individual, comprising administering to the individual an effective amount of a polypeptide that contains the Fc region variant. , to suppress the activation of B cells, mast cells, dendritic cells and/or basophils. In one embodiment, the "individual" is a human.

Another object of the invention is the use of a polypeptide containing a variant Fc region for the preparation or production of a drug. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the drug is designed to suppress the activation of B cells, mast cells, dendritic cells and/or basophils. In a further embodiment of the invention, the drug is for use in a method of suppressing the activation of B cells, mast cells, dendritic cells and/or basophils in an individual, comprising administering to the individual an effective amount of the drug to suppress the activation of B cells, mast cells, dendritic cells and/or basophils. In one embodiment, the "individual" is a human.

Another object of the invention is a method for suppressing the activation of B cells, mast cells, dendritic cells and/or basophils in an individual. In one embodiment, the method comprises administering to an individual an effective amount of a polypeptide comprising a variant Fc region to inhibit the activation of B cells, mast cells, dendritic cells, and/or basophils. In one embodiment, the "individual" is a human.

Another object of the invention are pharmaceutical compositions that contain a polypeptide containing a variant Fc region. In another embodiment of the invention, the pharmaceutical composition is designed to suppress the activation of B cells, mast cells, dendritic cells and/or basophils.

Polypeptides containing the Fc region variant of the present invention can suppress the activation of B cells, mast cells, dendritic cells and/or basophils. Without wishing to be bound by theory, this inhibition of activation is the result of the selective binding of inventive Fc region variants to Fc gamma RIIb without activation of the activating Fc gamma R. As used herein, "B cell activation" includes proliferation, production IgE, IgM production and IgA production. Without wishing to be bound by theory, it can be hypothesized that suppression of B cell activation results from the ability of the inventive Fc region variants in polypeptides containing the Fc region variant of the present invention to cross-link Fc-gamma RIIb with IgE to suppress production IgE B cells, with IgM to suppress IgM production by B cells, and with IgA to suppress IgA production. In addition to the above, inhibitory actions similar to those above result from direct or indirect cross-linking of Fc-gamma RIIb with molecules that are expressed on B cells and contain an ITAM domain within the cell, or interaction with an ITAM domain such as BCR, CD19 and CD79b. As used herein, "mast cell activation" includes proliferation, activation by IgE, and degranulation. Without wishing to be bound by theory, the suppression of mast cell activation is the result of the ability of the inventive Fc region variants in polypeptides containing the Fc region variant of the present invention to directly or indirectly crosslink Fc-gamma RIIb to IgE molecules. α-receptors that are expressed on mast cells and contain an ITAM domain, or to interact with an ITAM domain such as Fc-epsilon RI, DAP 12 and CD200R3. In the context of the present description, "activation of basophils" includes the proliferation and degranulation of basophils. Without wishing to be bound by theory, it can be hypothesized that suppression of basophil activation results from the ability of the inventive Fc region variants in polypeptides containing the Fc region variant of the present invention to directly or indirectly cross-link Fc-gamma RIIb to molecules on the cell wall. membrane that contain the ITAM domain within the cell, or to interact with the ITAM domain. In the context of the present description, "activation of dendritic cells" includes the proliferation and degranulation of dendritic cells. Without being bound by theory, it can be assumed that the suppression of dendritic cell activation results from the ability of the inventive Fc region variants in polypeptides containing the Fc region variant of the present invention to directly or indirectly crosslink Fc-gamma RIIb to molecules on cell membrane that contain the ITAM domain within the cell, or to interact with the ITAM domain.

Still further embodiments of the invention is a polypeptide comprising a variant Fc region as described herein for use in the treatment of an immunological inflammatory disease. In some embodiments, the invention provides a polypeptide comprising an Fc region variant for use in a method of treating an immunological inflammatory disease in a subject, comprising administering to the subject an effective amount of a polypeptide comprising an Fc region variant for the treatment of an immunological inflammatory disease. In one embodiment, the "individual" is a human.

The following object of the invention is the use of a polypeptide containing a variant Fc-region, presented in the present description, for the preparation or production of drugs. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the drug is for the treatment of an immunological inflammatory disease. In a further embodiment of the invention, the medicament is for use in a method for treating an immunological inflammatory disease in a subject, comprising administering to the subject an effective amount of a medicament for treating an immunological inflammatory disease. An "individual" in the context of any of the above embodiments of the invention may be a human.

A further aspect of the invention is a method for treating an immunological inflammatory disease in an individual. In one embodiment, the method comprises administering to an individual an effective amount of a polypeptide comprising a variant Fc region for the treatment of immunological inflammatory diseases. In one embodiment, the "individual" is a human.

The following object of the invention are pharmaceutical compositions containing any of the polypeptides containing the variant Fc region presented in the present description. In another embodiment of the invention, the pharmaceutical composition is for the treatment of an immunological inflammatory disease.

As described above, since the polypeptides containing the Fc region variant of the present invention can suppress the activation of B cells, mast cells, dendritic cells and/or basophils, the introduction of polypeptides containing the Fc region variant of the present invention, as a result, it can treat or prevent immunological inflammatory diseases.

In some embodiments, the immunological inflammatory disease to be treated is selected from the group consisting of rheumatoid arthritis, autoimmune hepatitis, autoimmune thyroiditis, autoimmune bullous diseases, autoimmune adrenocortical disease, autoimmune hemolytic anemia, autoimmune thrombocytopenic purpura, macrocytic neutrophic anemia, autoimmune autoimmune orchitis, autoimmune encephalomyelitis, autoimmune receptor disease, autoimmune infertility, chronic active hepatitis, glomerulonephritis, interstitial pulmonary fibrosis, multiple sclerosis, Paget's disease, osteoporosis, multiple myeloma, uveitis, acute and chronic spondylitis, gouty arthritis, inflammatory bowel disease, respiratory distress - adult syndrome (ARDS), psoriasis, Crohn's disease, Graves' disease, juvenile diabetes, Addison's disease, myasthenia gravis, phacoantigenic uveitis, systemic lupus erythematosus, allergic rhinitis, allergic dermatitis, ulcerative colitis, hypersensitivity, muscle degeneration, cachexia, systemic scleroderma, localized scleroderma, Sjögren's syndrome, Behcet's disease, Reiter's syndrome, type I and type II diabetes, impaired bone resorption, reaction graft-versus-host disease, ischemia-reperfusion injury, atherosclerosis, brain injury, cerebral malaria, sepsis, septic shock, toxic shock syndrome, fever, myalgia due to sedentary lifestyle (malgias due to staining), aplastic anemia, hemolytic anemia, idiopathic thrombocytopenia, Goodpasture's syndrome, Guillain-Barré syndrome, Hashimoto's thyroiditis, pemphigus, IgA nephropathy, hay fever, antiphospholipid antibody syndrome, polymyositis, Wegener's granulomatosis, nodular arteritis, mixed connective tissue disease, fibromyalgia, asthma, atopic dermatitis , chronic atrophic gastritis, primary biliary cirrhosis, primary sclerosing cholangitis, autoimmune pancreatitis, aortic inflammation syndrome, rapidly progressive glomerulonephritis, megaloblastic anemia, idiopathic thrombocytopenic purpura, primary hypothyroidism, idiopathic Addison's disease, insulin-dependent diabetes mellitus, chronic discoid lupus erythematosus, pemphigoid, herpes gravidis IgA, dependent bullous dermatosis, epidermolysis bullosa acquired, alopecia areata, vitiligo vulgaris, Sutton's centrifugal leukoderma, Harada's disease, autoimmune optic neuropathy, idiopathic azoospermia, recurrent miscarriage, hypoglycemia, chronic urticaria, ankylosing spondylitis, psoriatic arthritis, reactive arthritis, enteropathic, spondyloatropathy, enthesopathy, inflammatory bowel syndrome, chronic fatigue syndrome, dermatomyositis, myositis with inclusion bodies, Schmidt's syndrome, Graves' disease, pernicious anemia, lupoid hepatitis, in age-related dementia, Alzheimer's disease, demyelinating disorder, amyotrophic lateral sclerosis, hypoparathyroiditis, Dressler's syndrome, Eaton-Lambert syndrome, dermatitis herpetiformis, alopecia, progressive systemic sclerosis, CREST syndrome (calcinosis, Raynaud's syndrome, impaired esophageal motility, sclerodactyly and telangiectasia), sarcoidosis, rheumatic attack, erythema multiforme, Cushing's syndrome, transfusion reaction, Hansen's disease, Takayasu's arteritis, polymyalgia rheumatica, temporal arteritis, giant cell arthritis, eczema, lymphomatoid granulomatosis, Kawasaki disease, endocarditis, endomyocardial fibrosis, endophthalmitis, fetal erythroblastosis, eosinophilic fasciitis, Felty syndrome, Henoch-Schönlein purpura, transplant rejection, mumps, cardiomyopathy, purulent arthritis, familial Mediterranean fever, Muckle-Wells syndrome, and hyper-IgD syndrome.

Other embodiments of the invention relate to a polypeptide containing a variant Fc region, intended for the treatment or prevention of autoimmune diseases that may be caused or may be associated with the production of antibodies against self-antigens (autoantibodies). Some embodiments of the invention relate to a polypeptide containing an Fc region variant for use in a method for treating or preventing an autoimmune disease that may be caused by or may be associated with the production of antibodies against self antigens (autoantibodies) in an individual, comprising administering to the individual in an effective amount a polypeptide containing a variant Fc region for the treatment or prevention of an autoimmune disease. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

Another object of the invention is the use of a polypeptide containing a variant Fc region for the preparation or production of a drug. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the drug is for the treatment or prevention of an autoimmune disease which may be caused by or may be associated with the production of antibodies against self antigens (autoantibodies). In another embodiment, the medicament is for use in a method of treating or preventing an autoimmune disease that may be caused by or may be associated with the production of antibodies against self antigens (autoantibodies) in an individual, comprising administering to the individual an effective amount of a medicament for treating or preventing an autoimmune disease. diseases. An "individual" in the context of any of the above embodiments of the invention may be a human.

Another object of the invention is a method of treating or preventing an autoimmune disease which may be caused by or may be associated with the production of antibodies against self antigens (autoantibodies) in an individual. In one embodiment, the method includes administering to an individual an effective amount of a polypeptide comprising a variant Fc region to treat or prevent an autoimmune disease. In one embodiment, the "individual" is a human.

The following object of the invention are pharmaceutical compositions that contain any of the polypeptides containing the variant Fc region presented in the present description. Another embodiment of the invention is a pharmaceutical composition for the treatment or prevention of an autoimmune disease which may be caused by or associated with the production of antibodies against self antigens (autoantibodies).

Polypeptides containing the Fc region variant of the present invention can treat or prevent an autoimmune disease that may be caused by or may be associated with the production of antibodies against self antigens (autoantibodies) by suppressing the production of autoantibodies. The use of an antibody Fc-region fusion molecule with AchR (myasthenia gravis severe autoantigen) has been shown to inhibit the proliferation of B cells that express the AchR-recognizing BCR and induce apoptosis (J. Neuroimmunol 227, 2010, pp. 35-43). The use of a fusion protein formed by fusion of an Fc region variant of the present invention and an antigen recognized by an autoantibody provides cross-linking of Fc-gamma RIIb to the BCR of said autoantigen on a B cell, resulting in suppression of B cell proliferation and/or leads to the induction of B-cell apoptosis.

In some embodiments, the autoimmune disease that can be treated or prevented is selected from the group consisting of Guillain-Barré syndrome, pseudoparalytic myasthenia gravis, chronic atrophic gastritis, autoimmune hepatitis, primary biliary cirrhosis, primary sclerosing cholangitis, autoimmune pancreatitis, aortic inflammation syndrome, Goodpasture's syndrome, rapidly progressive glomerulonephritis, megaloblastic anemia, autoimmune hemolytic anemia, autoimmune neutropenia, idiopathic thrombocytopenic purpura, Graves' disease, Hashimoto's thyroiditis, primary hypothyroiditis, idiopathic Addison's disease, insulin-dependent diabetes mellitus, chronic discoid lupus erythematosus, pemphigoiderma, localized sclerosis herpes of pregnancy, linear IgA-dependent bullous dermatosis, acquired epidermolysis bullosa, alopecia areata, vitiligo vulgaris, Sutton's centrifugal acquired leukoderma, Harada's disease, autoimmune optic neuropathy, idiopathic azoospermia, recurrent miscarriage, type II diabetes, hypoglycemia, and chronic urticaria.

Further embodiments of the invention provide a polypeptide containing a variant Fc region for the treatment of a disease associated with a deficiency of a biologically important protein. In some embodiments, the invention is a polypeptide containing an Fc region variant for use in a method of treating a disease associated with a deficiency of a biologically important protein in an individual, comprising administering to the individual an effective amount of a polypeptide containing an Fc region variant to treat the disease. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

Another object of the invention is the use of a polypeptide containing a variant Fc region for the preparation or production of a drug. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the drug is for the treatment of a disease associated with a deficiency of a biologically important protein. In another embodiment, the drug is for use in a method of treating a disease associated with a deficiency of a biologically important protein in an individual, comprising administering to the individual an effective amount of a drug to treat the disease. An "individual" in the context of any of the above embodiments of the invention may be a human.

Another aspect of the invention is a method for treating a disease associated with a deficiency of a biologically important protein in an individual. In one embodiment, the method comprises administering to an individual an effective amount of a polypeptide containing a variant Fc region to treat a disease. In one embodiment, the "individual" is a human.

The following object of the invention are pharmaceutical compositions that contain any of the polypeptides containing the variant Fc region presented in the present description. In a further embodiment of the invention, the pharmaceutical composition is for the treatment of a disease associated with a deficiency of a biologically important protein.

In the case of diseases associated with a deficiency of a biologically important protein, therapeutic methods are used, in the implementation of which the protein is administered and added as a pharmaceutical agent. However, since the patient lacks protein from the very beginning, the protein added from the outside is recognized as a foreign substance and antibodies against the protein are produced. As a result, the protein is easily removed and the pharmaceutical effect is reduced. The use of a fusion protein containing this protein and an Fc region variant of the present invention cross-links Fc-gamma RIIb to a BCR that recognizes the protein on B cells, resulting in suppression of antibody production against the protein.

In some embodiments, the protein to be added is selected from the group consisting of factor VIII, factor IX, TPO, EPO, alpha-iduronidase, iduronate sulfatase, A-type heparan-N-sulfatase, B-type alpha-N-acetylglucosaminidase, acetylCoA: C-type alpha-glucosaminide acetyltransferase, D-type N-acetylglucosamine-6-sulfatase, galactose-6-sulfatase, N-acetylgalactosamine-4-sulfatase, beta-glucuronidase, alpha-galactosidase, acid alpha-galactosidase and glucocerebrosidase. These proteins can be added in diseases such as hemophilia, idiopathic thrombocytopenic purpura, renal anemia and lysosomal disease (mucopolysaccharidosis, Fabry disease, Pompe disease and Gaucher disease) (but not limited to them).

Further embodiments of the invention is a polypeptide containing a variant Fc region for use in the treatment of a viral infection. In some embodiments, the invention is a polypeptide containing a variant Fc region for use in a method of treating a viral infection in an individual, comprising administering to the individual an effective amount of a polypeptide containing a variant Fc region to treat a viral infection. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

The following object of the invention is the use of a polypeptide containing a variant Fc-region, presented in the present description, for the preparation or production of drugs. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In a further embodiment of the invention, the drug is for the treatment of a viral infection. In another embodiment, the drug is for use in a method for treating a viral infection in a subject, comprising administering to the subject an effective amount of a drug for treating a viral infection. An "individual" in the context of any of the above embodiments of the invention may be a human.

The next object of the invention is a method of treating a viral infection in an individual. In one embodiment, the method comprises administering to an individual an effective amount of a polypeptide comprising a variant Fc region to treat a viral infection. In one embodiment, the "individual" is a human.

The following object of the invention are pharmaceutical compositions that contain any of the polypeptides containing the variant Fc region presented in the present description. In a further embodiment of the invention, the pharmaceutical composition is for the treatment of a viral infection.

Antiviral antibodies that contain the Fc region variant of the present invention can suppress antibody-dependent enhancement (infections) characteristic of canonical antiviral antibodies. Antibody-dependent enhancement is a phenomenon in which a virus bound to an antibody undergoes phagocytosis by activating Fc-gamma R, whereby virus infection of the cell is enhanced. The binding of anti-Dengue antibodies to Fc-gamma RIIb has been shown to play an important role in suppressing antibody-dependent enhancement (Proc. Natl. Acad. Sci. USA 108, 2011, pp. 12479-12484). Cross-linking of Fc-gamma RIIb molecules with an immune complex comprising anti-dengue antibodies and Dengue virus inhibits Fc-gamma R mediated phagocytosis, resulting in suppression of antibody-dependent enhancement. Examples of these viruses include, but are not limited to, Dengue virus (DENV1, DENV2, DENV3 and DENV4) and HIV.

Further embodiments of the invention is a polypeptide containing a variant Fc region for use in the prevention or treatment of arteriosclerosis. In some embodiments, the polypeptide comprising the Fc region variant is for use in a method for preventing or treating arteriosclerosis in a subject, comprising administering to the subject an effective amount of the polypeptide comprising the Fc region variant to prevent or treat arteriosclerosis. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

Another object of the invention is the use of a polypeptide containing a variant Fc region for the preparation or production of a drug. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the drug is for the prevention or treatment of arteriosclerosis. In a further embodiment of the invention, the medicament is for use in a method for preventing or treating arteriosclerosis in a subject, comprising administering to the subject an effective amount of a medicament for preventing or treating arteriosclerosis. An "individual" in the context of any of the above embodiments of the invention may be a human.

The following object of the invention is a method for the prevention or treatment of arteriosclerosis in an individual. In one embodiment, the method comprises administering to an individual an effective amount of a polypeptide comprising a variant Fc region to prevent or treat arteriosclerosis. In one embodiment, the "individual" is a human.

The following object of the invention are pharmaceutical compositions that contain any of the polypeptides containing the variant Fc region presented in the present description. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the pharmaceutical composition is for the prevention or treatment of arteriosclerosis.

Antibodies against oxidized LDL, i.e. causative factor of arteriosclerosis containing the variant Fc region of the present invention can prevent Fc-gamma RIIa dependent inflammatory cell adhesion. Although antibodies against oxidized LDL inhibit the interaction between oxidized LDL and CD36, antibodies against oxidized LDL bind to endothelial cells and monocytes recognize their Fc region in an Fc-gamma RIIa-dependent or Fc-gamma RI-dependent manner (Immunol. Lett. 108, 2007, pp. 52-61). The use of antibodies containing the Fc region variant of the present invention can inhibit Fc-gamma RIIa-dependent binding and inhibit monocyte adhesion via Fc-gamma mediated inhibitory signals.

Another embodiment of the invention is a polypeptide containing a variant Fc region for use in the prevention or treatment of cancer. In some embodiments, the Fc region variant-containing polypeptide is for use in a method for preventing or treating cancer in a subject, comprising administering to the subject an effective amount of the Fc region variant-containing polypeptide to prevent or treat cancer. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

The next object is the use of a polypeptide containing a variant Fc region, presented in the present description, for the preparation or production of drugs. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment, the drug is for the prevention or treatment of cancer. In a further embodiment of the invention, the medicament is for use in a method for preventing or treating cancer in a subject, comprising administering to the subject an effective amount of a medicament for preventing or treating cancer. An "individual" in the context of any of the above embodiments of the invention may be a human.

The next object of the invention is a method for the prevention or treatment of cancer in an individual. In one embodiment, the method includes administering to an individual an effective amount of a polypeptide containing a variant Fc region to prevent or treat cancer. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In one embodiment of the invention, the "individual" is a human.

The following object of the invention are pharmaceutical compositions that contain any of the polypeptides containing the variant Fc region presented in the present description. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the pharmaceutical composition is for the prevention or treatment of cancer.

As described above, enhancing Fc-gamma binding of RIIb is known to increase the agonist activity of an agonist antibody and enhance the antitumor activity of the antibody. Thus, agonist antibodies containing the Fc region variant of the present invention can be used to treat or prevent cancer. In particular, the Fc region variant of the present invention enhances the agonistic activity of antibodies directed against, for example, TNF receptor family receptors such as CD120a, CD120b, lymphotoxin receptor beta, CD134, CD40, FAS, TNFRSF6B, CD27, CD30 , CD137, TNFRSF10A, TNFRSF10B, TNFRSF10C, TNFRSF10D, RANK, osteoprotegerin, TNFRSF12A, TNFRSF13B, TNFRSF13C, TNFRSF14, nerve growth factor receptor, TNFRSF17, TNFRSF18, TNFRSF19, TNFRSF21, TNFRSF18, TNFRSF19, TNFRSF21, TNFRSF18, or ectodysplasin A receptor can be used to prevent and treat them, and ectodysplasin A 25 cancer. In addition, the agonistic activity of antibodies against other molecules that have agonistic activity as a result of interaction with Fc-gamma RIIb is also increased. In addition, by incorporating a variant of the Fc region of the present invention into an anti-receptor tyrosine kinase (RTK) antibody such as Kit that inhibits cell proliferation by cross-linking with Fc-gamma RIIb, the inhibitory effect of anti-cell proliferation antibodies can be enhanced.

In some embodiments, the invention provides a method for the prevention or treatment of cancer, including (but not limited to) a type of cancer selected from: small cell lung cancer, non-small cell lung cancer, pulmonary adenocarcinoma and squamous cell lung carcinoma, large intestinal cancer, rectal cancer, cancer colon, breast cancer, gastric cancer, pancreatic cancer, kidney cancer, prostate cancer, ovarian cancer, thyroid cancer, cholangiocarcinoma, peritoneal cancer, mesothelioma, squamous cell carcinoma, cervical cancer, endometrial cancer, bladder cancer, cancer esophagus, head and neck cancer, nasopharyngeal cancer, salivary gland cancer, thymoma, skin cancer, basal cell carcinoma, malignant melanoma, anus cancer, penile cancer, testicular cancer, Wilms tumor, acute myeloid leukemia (including acute myeloid leukemia, acute myeloid leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia and chronic myelogenous leukemia, acute lymphoblastic leukemia, chronic lymphatic leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma (Burkitt's lymphoma, chronic lymphocytic leukemia, mucosal fugoid, mantle cell lymphoma, follicular lymphoma, diffuse large cell lymphoma, piloid leukemoid plasmacytoma, peripheral T-cell lymphoma and adult T-cell leukemia/T-cell lymphoma), Langerhans cell histocytosis, multiple myeloma, myelodysplastic syndrome, brain tumors (including glioma, astroglioma, glioblastoma, meningioma and ependymoma), neuroblastoma , retinoblastoma, osteosarcoma, Kaposi's sarcoma, Ewing's sarcoma, angiosarcoma and hemangiopericytoma.

Antibodies that have been modified to increase binding activity to Fc-gamma R, including Fc-gamma RIIb, by modifying at least one amino acid residue, may enhance antigen clearance from plasma, as described or suggested, for example, in WO 2013/047752, WO 2013/125667, WO 2014/030728 or WO 2014/163101.

Without wishing to be bound by a particular theory, an antibody having increased Fc-gamma R-binding activity under neutral pH conditions rapidly enters cells along with the antigen complexed with the antibody, and as a result, elimination of the antigen from plasma can be enhanced when the antibody is administered in vivo.

Antibodies that have been modified to have an increased pI, by modifying at least one amino acid residue that can be displayed on the surface of the antibody, may be more rapidly incorporated into cells, or may enhance antigen clearance from plasma, as described or suggested, for example in WO 2007/114319, WO 2009/041643, WO 2014/145159 or WO 2012/016227.

Without going into a particular theory, it can be assumed that the pH of biological fluids (eg plasma) is in the range of neutral pH values. In biological fluids, the net positive charge of an antibody with increased pI increases due to the increase in pI, and as a result, the antibody is attracted with greater force through physicochemical Coulomb interaction to the endothelial cell surface, which has a net negative charge, compared to an antibody that does not have an increased pI. . This means that through such non-specific binding, cell penetration of the antibody in combination with its antigen is increased, resulting in increased elimination of the antigen from the plasma. It is assumed that this phenomenon is widespread in vivo regardless of cell type, tissue type, organ type, etc.

In the context of the present description, increased elimination of an antigen from plasma means, for example, that the rate of elimination of an antigen from plasma is increased compared to the rate when a polypeptide is administered that contains the corresponding but not similarly modified Fc region. The increase in antigen elimination from plasma can be assessed, for example, by measuring the plasma concentration of the antigen. Various methods are known in the art for measuring plasma antigen concentration.

In other embodiments, the invention is a polypeptide containing a variant Fc region for use in enhancing antigen clearance from plasma. In some embodiments, the invention is a polypeptide that contains an Fc region variant for use in a method of enhancing elimination of an antigen from plasma in an individual, comprising administering to the individual an effective amount of a polypeptide that contains an Fc region variant to enhance elimination of the antigen. from plasma. In these embodiments, it is preferred that the polypeptide further comprises an antigen-binding domain. "Individual" in the context of any of the above embodiments of the invention is preferably a human.

Another object of the invention is the use of a polypeptide that contains a variant Fc region for the manufacture or production of a drug. In some aspects of the invention, the polypeptide is an antibody. In some aspects of the invention, the polypeptide is an Fc-containing fusion protein. In another embodiment of the invention, the drug is intended to enhance the elimination of antigen from plasma. In a further embodiment of the invention, the drug is for use in a method for enhancing plasma antigen clearance in an individual, comprising administering to the subject an effective amount of the drug to enhance plasma antigen clearance. In these embodiments, it is preferred that the polypeptide further comprises an antigen-binding domain. An "individual" in the context of any of the above embodiments of the invention may be a human.

Another embodiment of the invention is a method for enhancing the elimination of an antigen from plasma in an individual. In one embodiment of the invention, the method comprises administering to an individual an effective amount of a polypeptide that contains a variant Fc region to enhance clearance of the antigen from plasma. In these embodiments, it is preferred that the polypeptide further comprises an antigen-binding domain. In one embodiment, the "individual" is a human.

The following object of the invention are pharmaceutical compositions that contain any of the polypeptides containing the variant Fc region presented in the present description. The next embodiment of the invention is a pharmaceutical composition designed to enhance the elimination of antigen from plasma. In this embodiment, it is preferred that the polypeptide further comprises an antigen binding domain.

In one of the embodiments of the invention, compared with the variant before the amino acid modification, an antibody that has an Fc region variant of the present invention increases the elimination of antigen from plasma, for example, at least 1.1, 1.25, 1.5 , 1.75, 2.0, 2.25, 2.5, 2.75, 3.0, 3.25, 3.5, 3.75, 4.0, 4.25, 4.5, 4 .75, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10.0 times or more at administration of the antibody in vivo.

A further aspect of the invention are methods of preparing a medicament or pharmaceutical composition comprising mixing any polypeptides containing Fc region variants provided herein with a pharmaceutically acceptable carrier, for example, for use in any of the above therapeutic methods. In one of the embodiments of the invention, methods for preparing a drug or pharmaceutical composition further include adding at least one additional therapeutic agent to the drug or pharmaceutical composition.

The antibodies of the invention can be used for treatment either alone or in combination with other agents. For example, an antibody of the invention may be co-administered with at least one additional therapeutic agent.

Similarly, polypeptides containing a variant Fc region of the invention can be used for treatment either alone or in combination with other agents. For example, a polypeptide containing a variant Fc region of the invention may be co-administered with at least one additional therapeutic agent.

Such combination therapies as mentioned above include co-administration (when two or more therapeutic agents are included in the same or different compositions) and separate administration, in each case administration of an antibody or polypeptide containing an Fc region variant of the invention, may be performed before, concurrently and/or after administration of the additional therapeutic agent or agents. In one embodiment, administration of the anti-myostatin antibody and administration of the additional therapeutic agent may be within about a month, or within about 1, 2, or 3 weeks, or within about 1, 2, 3, 4, 5, or 6 days of each other. friend.

In another embodiment, the administration of the polypeptide containing the Fc region variant and the administration of the additional therapeutic agent can be within about a month, or within about 1, 2, or 3 weeks, or within about 1, 2, 3, 4, 5, or 6 days relative to each other.

An antibody or polypeptide containing an Fc region variant of the invention (and optionally any additional therapeutic agent) can be administered by any suitable route, including parenteral, intrapulmonary and intranasal, and, if necessary, used for topical treatment, injection into damage. Parenteral infusions include intramuscular, intravenous, intra-arterial, intraperitoneal or subcutaneous administration. Dosing may be by any suitable route, for example, by injection, such as intravenous or subcutaneous injection, depending, inter alia, on whether the treatment is short-term or chronic. Various dosing regimens may be used, including (but not limited to) single administration or multiple administrations at different time points, bolus administration, and pulsatile infusion.

Antibodies or polypeptides containing the Fc region variant of the invention may be formulated, dosed, and administered in accordance with good clinical practice. Factors considered in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of administration of the agent, the method of administration, the schedule of administration, and other factors known to medical practitioners. The antibody may, but is not required, be included in the composition in combination with one or more other agents currently used to prevent and treat the disorder in question. The effective amount of these other agents depends on the amount of antibody present in the composition, the type of disorder or treatment, and other factors mentioned above. They are generally used in the same doses and using the same routes of administration as specified in the present description, or in an amount ranging from 1 to 99% of the dose indicated in the present description, or in any dose and with any pathways that are empirically/clinically judged to be suitable.

For the prevention or treatment of a disease, the acceptable dose of an antibody of the invention (when used alone or in combination with one or more other additional therapeutic agents) should depend on the type of disease being treated, the type of antibody, the type of polypeptide containing the variant Fc region , severity and course of disease, whether the antibody or polypeptide containing the Fc region variant is used for preventive or therapeutic purposes, prior therapy, the patient's medical history and response to the antibody or polypeptide containing the Fc region variant, as well as the prescription of the attending physician. An antibody or polypeptide containing a variant Fc region can be administered to a patient at a single time or as a series of treatments. Depending on the type and severity of the disease, from about 1 μg/kg to 15 mg/kg (e.g., 0.5-10 mg/kg) of the antibody may represent the initial possible dose to be administered to the patient, e.g., via one or more separate treatments. or by continuous infusion. One example of a typical daily dose may be from about 1 μg/kg to 100 mg/kg or more, depending on the above factors. When repeated administrations are used over a period of several days or a longer period of time, depending on the condition, treatment should generally be continued until suppression of the symptoms of the disease is achieved. One example of doses of an antibody or polypeptide containing a variant Fc region is from about 0.05 to about 10 mg/kg. Thus, one or more of the following doses may be administered to a patient: about 0.5, 2.0, 4.0, or 10 mg/kg (or any combination thereof). These doses can be administered in fractions, such as every week or every three weeks (eg, so that the patient receives from about two to about twenty, for example, about six doses of an antibody or polypeptide containing a variant Fc region). An initial increased loading dose may be used followed by one or more lower doses. Using conventional methods and assays, the effect of said therapy can be easily monitored.

Obviously, any of the above compositions or therapeutic methods can be used using the immunoconjugate of the invention, instead of or in addition to the antibody to myostatin.

Obviously, any of the above compositions or therapeutic methods can be used using the immunoconjugate of the invention, instead of or in addition to the polypeptide containing the variant Fc region presented in the present description.

3. Products

Another object of the invention is a product that contains products used for the treatment, prevention and/or diagnosis of the above disorders. The product is a container and a label or package insert placed on or inside the container. Acceptable containers are, for example, jars, vials, syringes, intravenous solution bags, etc. Containers can be made from various materials such as glass or plastic. The container contains a composition that alone or in combination with another composition is effective in treating, preventing, and/or diagnosing a condition, and may have a sterile access port (for example, the container may be an intravenous solution bag or vial provided with a stopper that can be pierced using a hypodermic needle). At least one active ingredient in the composition is an antibody of the invention. The label or package insert indicates that the composition is used to treat the selected condition. In addition, the product may include (a) a first container containing the composition, where the composition contains the antibody proposed in the present invention; and (b) a second container containing the composition, wherein the composition contains an additional cytotoxic or other therapeutic agent. According to this embodiment of the invention, the product may contain a package insert leaflet that contains information that the compositions can be used to treat a particular condition. Alternatively or additionally, the article of manufacture may further comprise a second (or third) container with a pharmaceutically acceptable buffer such as bacteriostatic water for injection (BVI), phosphate buffered saline, Ringer's solution and dextrose solution. In addition, it may include other products required from a commercial and consumer point of view, in particular other buffers, diluents, filters, needles and syringes.

As should be obvious, any of the above products may include the immunoconjugate of the invention, instead of or in addition to the antibody to myostatin.

Examples

Below are examples of methods and compositions proposed in the invention. As should be apparent, various other embodiments of the invention may be practiced in view of the general description of the invention provided above.

Example 1

Expression and purification of latent and mature forms of human myostatin, cynomolgus monkeys and mice

Human latent myostatin (herein also referred to as the latent form of human myostatin) (SEQ ID NO: 1) was transiently expressed using FreeStyle293-F cells (FS293-F cells) (Thermo Fisher, Carlsbad, CA, USA) . Conditioned media containing the expressed latent form of human myostatin were acidified to pH 6.8 and diluted 1/2 volume with milliQ water before being loaded onto a Q-sepharose FF ion exchange column (GE healthcare, Uppsala, Sweden). The pH of the permeate fraction was adjusted to pH 5.0 and loaded onto a HP SP Sepharose cation exchange column (GE healthcare, Uppsala, Sweden) and then eluted using a NaCl gradient. Fractions containing latent human myostatin were collected and then subjected to gel filtration on a Superdex 200 column (GE healthcare, Uppsala, Sweden) equilibrated with 1×PBS. Then the fractions containing the latent form of human myostatin were pooled and stored at -80°C.

Human mature myostatin (herein also referred to as mature human myostatin) (SEQ ID NO: 2) was purified from purified latent human myostatin. Latent human myostatin was acidified with 0.1% trifluoroacetic acid (TFA) and applied to a Vydac 214TP C4 reverse phase column (Grace, Deerfield, IL, USA) and eluted using a TFA/CH ₃ CN gradient. Fractions containing human mature myostatin were pooled, dried and stored at -80°C. For reconstitution, human mature myostatin was dissolved in 4 mM HCl.

Expression and purification of latent and mature myostatin from cynomolgus monkeys (cynomolgus or cyno) (SEQ ID NOS: 3 and 4, respectively) and mice (SEQ ID NOS: 5 and 6, respectively) was carried out in all cases in exactly the same manner as in the case of human copy. With respect to sequence homology, mature human, cyno, and mouse myostatin were 100% identical, so any mature myostatin, regardless of species, was used as mature myostatin in all experiments performed.

Example 2

Identification of antibodies to latent myostatin

Antibodies to latent myostatin were obtained, selected and analyzed as follows.

New Zealand White Rabbits (NZW) aged 12-16 weeks were immunized intradermally with mouse latent myostatin and/or human latent myostatin (50-100 μg/dose/rabbit). Treatment with the indicated dose was repeated 3-4 times within 1 month. Spleens and blood samples from the immunized rabbit were collected 1 week after the last immunization. Antigen-specific B cells were stained with labeled antigen, sorted using an FCM cell sorter (FACS aria III, BD) and plated in 96-well plates at a density of 1 cell/well along with 25,000 EL4 cells/well (cells from the European Cellular Collection). cultures) and together with conditioned medium for rabbit T-cells, diluted 20 times, and cultured for 7-12 days. EL4 cells were pretreated with mitomycin C (Sigma) for 2 hours and washed 3 times. Rabbit T cell conditioned medium was prepared by culturing rabbit thymocytes in RPMI-1640 medium containing phytohemagglutinin-M (Roche), phorbol-12-myristate-13-acetate (Sigma) and 2% FBS. After cultivation, the supernatants of B-cell cultures were collected for additional analysis and the debris was cryopreserved.

An ELISA assay was used to evaluate the specificity of antibodies in the B cell culture supernatant. MAXISorp 384-well plates (Nunc) were sensitized with streptavidin (GeneScript) at 50 nM in PBS for 1 hour at room temperature. The plates were then blocked with Blocking One reagent (Nacalai Tesque) diluted 5 times. Human or mouse latent myostatin was labeled with NHS-PEG4-biotin (PIERCE) and added to blocked ELISA plates, incubated for 1 hour and washed with Tris buffered saline with 0.05% Tween-20 (TBS-T). B cell culture supernatants were loaded onto ELISA plates, incubated for 1 h and washed with TBS-T. Binding was detected with goat anti-rabbit IgG conjugated to horseradish peroxidase (BETHYL) with ABTS (KPL).

A total of 17818 B cell lines were screened for specific binding to mouse and/or human latent myostatin, 299 lines were selected and designated as MST0255-287, 630-632, 677-759, 910, 932-1048, 1050-1055 , 1057-1066, 1068, 1070-1073, 1075-1110, 1113-1119. RNA was purified from the respective cell debris using ZR-96 Quick-RNA kits (ZYMO RESEARCH).

DNA encoding the heavy and light chain variable regions of each selected cell line was amplified by reverse transcription PCR and cloned into expression vectors with the Glm heavy chain constant region sequence (SEQ ID NO: 50 (amino acid sequence shown in SEQ ID NO: 7) ) and with the light chain constant region sequence k0MTC or k0MC (SEQ ID NO: 53 or 194 (amino acid sequence (same in both cases) shown in SEQ ID NO: 8)) respectively. Recombinant antibodies were transiently expressed using FreeStyle FS293-F and 293fectin cells (Life technologies) according to the manufacturer's instructions. Culture supernatant or recombinant antibodies were used for screening. Recombinant antibodies were purified with protein A (GE Healthcare) and eluted in D-PBS or His buffer (20 mM histidine, 150 mM NaCl, pH 6.0). Additionally, if necessary, gel filtration was performed to remove the high molecular weight and/or low molecular weight component.

Example 3

Characterization of Anti-Latent Myostatin Antibody (HEK-Blue-Assay (BMP1-Activation))

Reporter gene analysis was used to assess the biological activity of active myostatin in vitro. HEK-Blue™ TGF-beta cells (Invivogen), which express Smad3/4-binding element (SBE)-indicated SEAP (Secreted Alkaline Phosphatase) reporter genes, allow for the detection of bioactive myostatin by monitoring activation of activin type 1 and type 1 activin receptors. 2. Active myostatin stimulates the production of SEAP, which is secreted into the cell supernatant. The amount of SEAP secreted is then determined using QUANTIBlue™ (Invivogen).

HEK-Blue™ TGF-beta cells were maintained in DMEM (Gibco) supplemented with 10% fetal bovine serum, 50 µg/ml streptomycin, 50 U/ml penicillin, 100 µg/ml Normocin™, 30 µg/ml blasticidin, 200 µg/ml HygroGold™ and 100 µg/ml Zeocin™. When performing a functional assay, cell medium was changed to assay medium (DMEM supplemented with 0.1% bovine serum albumin, streptomycin, penicillin, and Normocin™) and plated in a 96-well plate. Patent human, cynomolgus or mouse myostatin was incubated with recombinant human BMP1 (R&D Systems) and latent myostatin antibody at 37° C. overnight. Sample mixtures were transferred into cells. After a 24 hour incubation, cell supernatants were mixed with QUANTIBlue™ and absorbance measured at 620 nm in a colorimetric plate reader. As shown in FIG. 1, mAb MST1032-G1m (see Table 2a for amino acid and nucleotide sequence) interfered with protease-mediated activation of latent human, cynomolgus, and mice myostatin, as evidenced by reduced concentrations of secreted SEAP. It was demonstrated that MST1032-G1m had almost comparable inhibitory activity against human latent myostatin, cynomolgus and mice.

Example 4

Characterization of Anti-Latent Myostatin Antibody (HEK Blue Assay (Spontaneous Activation))

Human, cynomolgus or mouse latent myostatin was incubated with anti-latent myostatin antibody at 37° C. overnight and the sample mixture was loaded into HEK-Blue™ TGF-beta cells. After a 24 hour incubation, the cell supernatant was mixed with QUANTIBlue™ and the absorbance was measured at 620 nm in a colorimetric plate reader. As shown in FIG. 2, after incubation at 37° C., active myostatin was released from the latent form. In the presence of mAb MST1032-G1m, myostatin activation was inhibited and, as a result, a lower level of SEAP was detected in the cell supernatant. MST1032-G1m inhibited spontaneous activation of latent myostatin and showed almost comparable inhibitory activity against human, cynomolgus and mice latent myostatin.

Example 5

Characterization of anti-latent myostatin antibody (ELISA)

Unsensitized ELISA plates (NUNC-IMMUNO plate, MAXISORP surface, Nalge Nunc International) were coated with 20 μl of 50 nM streptavidin (GenScript) for 1 hour at room temperature. The plates were then washed three times with PBS and blocked overnight with 50 μl of 20% Blocking One (Nacalai Tesque). The following day, each well of each plate was incubated with biotinylated mature myostatin, human or mouse biotinylated latent myostatin, or biotinylated recombinant mouse myostatin propeptide-Fc chimeric protein (R&D Systems) at 4 nM/well/20 μl for 2 hours. After washing 20 μl of antibody sample was added to the wells and the plates were incubated for 1 hour. The plates were washed and 20 μl of anti-human IgG conjugated with horseradish peroxidase (HRP) (Abeam) diluted in HEPES buffered saline was added and the plates were incubated for another 1 hour. The plates were then washed again and then 50 μl of ABTS (KPL) were added to each well and the plates were incubated for 1 hour. The signal was detected at 405 nm in a colorimetric plate reader. The results of the binding experiment are shown in FIG. 3. MST1032-G1m binds to latent myostatin (ie, a non-covalent complex of mature myostatin and propeptides) and propeptides, but does not bind to mature myostatin. These results indicate that MST1032-G1m specifically binds to myostatin propeptide (eg, propeptide portion) but does not bind to active myostatin (eg, mature region).

Example 6

Characterization of antibodies to latent myostatin (Western blotting)

Mouse latent myostatin was incubated with recombinant human BMP1 (R&D Systems) with MST1032-G1m or no antibody at 37° C. overnight. The samples were then mixed with 4x SDS-PAGE reconstitution sample buffer (Wako) and kept at 95° C. for 5 minutes and then loaded onto the SDS gel electrophoresis system. Proteins were transferred to the membrane using a Trans-Blot Turbo™ transfer system (Bio-rad). Myostatin propeptide was detected using a sheep anti-mouse GDF8 propeptide antibody (R&D Systems), which was then detected using an anti-sheep IgG-HRP antibody (Santa Cruz). The membrane was incubated with ECL substrate and visualized using an ImageQuant LAS 4000 (GE Healthcare). As shown in FIG. 4, propeptide cleavage by BMP1 was inhibited by MST1032-G1m.

Example 7

Characterization of Anti-Latent Myostatin Antibody (BIACORE®)

The kinetic parameters of antibodies to latent myostatin against human latent myostatin, cynomolgus monkeys (cyno) and mice were evaluated at 37° C. at pH 7.4 using a BIACORE® T200 device (GE Healthcare). ProA/G (Protein A/G) (Pierce) was immobilized on all flow cells of the CM4 chip using an amine coupling kit (GE Healthcare). Anti-latent myostatin antibody and analytes were prepared in ACES, pH 7.4 (20 mM ACES, 150 mM NaCl, 1.2 mM CaCl ₂ , 0.05% Tween 20, 0.005% NaN ₃ ). The antibody was "captured" on the surface of the sensor chip using ProA/G. Antibody immobilization levels typically corresponded to 150-220 resonance units (RU). Human, cyno, or mouse latent myostatin was then injected at concentrations ranging from 3.125 to 50 nM, which were obtained by two-fold serial dilutions and then allowed to dissociate. The sensor surface was regenerated with 25 mM NaOH. Kinetic parameters were determined by processing and fitting the data to a 1:1 binding model using BIACORE® T200 Evaluation software version 2.0 (GE Healthcare). The sensorgrams are shown in Fig. 5. Association rate (ka), dissociation rate (kd), and binding affinity (KD) data are shown in Table 3. MST1032-G1m kinetic parameters characterizing binding to latent human, cyno, and mouse myostatin were comparable.

Example 8

In vivo efficacy of anti-latent myostatin antibody on muscle mass and fat mass

The in vivo efficacy of the mAb MST1032-G1m was studied in mice. In this experiment, the antibody to mature myostatin 41C1E4 (described in US No. 7632499) was used as a positive control. To avoid potential immunomodulation due to the response in mice to human antibody, in vivo experiments were performed in mice with severe combined immunodeficiency (SCID). SCID (C.B-17 SCID) mice aged 5 weeks (Charles River Laboratories Japan, Inc. (Kanagawa, Japan)) were treated with monoclonal antibodies at various doses or vehicle (PBS), which were administered intravenously once a week for 2 weeks. On days 0, 7 and 14, the total lean mass and fat mass of the mice were determined by nuclear magnetic resonance (NMR) (minispec LF-50 device, Bruker Bio Spin, (Kanagawa, Japan)). Animals were euthanized on day 14 and the gastrocnemius and quadriceps muscles were removed and weighed.

Statistical significance was determined by analysis of variance, Student's t-test and Dunnett's test using JMP 9 software (SAS, Inc.). A p value below 0.05 was considered significant.

The results of this experiment are shown in Fig. 6A-B. Both antibodies (MST1032-G1m and 41C1E4) increased lean body mass in a dose-dependent manner, and MST1032-G1m, when administered at 40 mg/kg, significantly reduced fat mass compared to the vehicle-treated (PBS) group at day 14.

After two weeks of treatment, the antibodies, which were administered at doses of 10 and 40 mg/kg, resulted in a significant increase in wet weight of the quadriceps and gastrocnemius muscles relative to the vehicle-treated group.

Example 9

Comparison in vivo of various related anti-myostatin antibodies

All experimental schemes used were as described in Example 8. Various related antibodies to myostatin 41C1E4, REGN, OGD, and MYO-029 were tested in 5 week old SCID mice for 2 weeks. REGN, OGD and MYO-029 are anti-myostatin antibodies described in WO 2011/150008 as H4H1657N2, in WO 2013/186719 as OGD1.0.0. and in WO 2004/037861 as MYO-029, respectively. These antibodies were administered intravenously once a week. Lean mass and body fat mass were determined by whole body NMR scanning on days 0 and 14. Compression force was measured with a compression force test device (e.g., GPM-100B, MELQUEST Ltd., (Toyama, Japan)) per day 14. As demonstrated in the results presented in FIG. 7A-B, lean body mass was increased with these antibodies other than MYO-029 (P>0.05). The grip force in the MST1032-G1m treated group was significantly increased compared to the vehicle-treated (PBS) group when applied at a dose of 10 mg/kg (P<0.05). 41C1E4 and REGN also significantly increased compressive strength compared to filler treatment (PBS). Total body fat mass tended to decrease with MST1032-G1m at 10 mg/kg. MST1032-G1m was more effective in reducing fat mass than other antibodies to mature myostatin.

Example 10

Humanization of antibodies to latent myostatin

Variable region amino acid residues were numbered according to Kabot (Kabat et al., 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, 1991).

Heavy and light chain variable regions were created for humanized MST1032 antibodies. Some of the humanized MST1032 antibodies contained a reverse mutation in the framework region. The polynucleotides of the designed heavy and light chain variable regions were synthesized by GenScript Inc. and cloned into expression vectors containing the SGI heavy chain constant region sequence (SEQ ID NO: 52 (amino acid sequence shown in SEQ ID NO: 9)) and light chain constant region sequence SKI (SEQ ID NO: 54 (amino acid sequence shown in SEQ ID NO: 10)) respectively. Humanized antibodies were transiently expressed in FS293-F cells and HEK Blue and BIACORE® assays were performed as described above. As shown in FIG. 8 and in Table 4, and when compared with FIG. 1 and 2, the humanized antibody (MS1032LO00-SG1) was found to have comparable inhibitory activity and affinity to the chimeric antibody (MST1032-G1m).

Example 11

Creation of pH-Dependent Antibody to Latent Myostatin To generate pH-dependent antibodies to latent myostatin, histidine-scanning mutagenesis of all CDRs of mAb MS1032LO00-SG1 was performed. Each amino acid in the CDR was individually replaced with histidine using an In-Fusion HD cloning kit (Clontech Inc. or Takara Bio company) according to the manufacturer's instructions. After sequencing confirmed the correct mutation in each variant, the variants were transiently expressed and purified as described above. All histidine substitution variants were evaluated using the BIACORE® assay modified from the assay described above. In general, the method was as follows: in the BIACORE® assay, an additional dissociation phase at pH 5.8 was carried out immediately after the dissociation phase at pH 7.4. This was done to evaluate the pH-dependent dissociation of antibody (Ab) and antigen (Ag) from complexes prepared at pH 7.4 compared to the corresponding dissociation at pH 5.8. The dissociation rate in pH 5.8 buffer was determined by data processing and fitting using Scrubber 2.0 curve fitting software (BioLogic Software).

As shown in FIG. 9, the parent antibody (Ab001) showed no decrease in binding response at pH 5.8 compared to the dissociation phase at pH 7.4. Several single histidine substitutions resulted in a moderate to strong reduction in binding response at pH 5.8 compared to the dissociation phase at pH 7.4. The dissociation rate at pH 5.8 for each variant with a single histidine substitution is presented in Table 5. As shown in Table 5, Ab002 exhibited the fastest dissociation rate from the Ab/Ag complex at pH 5.8, exceeding more than 200 times the rate for the parent antibody (Ab001). Then created antibodies with a combination of these mutations in the CDR. The CDR sequences of antibodies containing these various mutations are shown in Table 6.

Example 12

Increasing the affinity of a pH-dependent antibody to latent myostatin

To identify mutations that increase affinity at pH 7.4 and/or in vitro inhibitory activity, more than 500 heavy and light chain variants, respectively, were generated using at least one variant generated in Example 11 as a template. In these embodiments, each amino acid in the CDR was replaced with 18 other amino acids, with the exception of the original amino acid and cysteine. The binding capacity of the variants to human latent myostatin was evaluated at 37° C. at pH 7.4 using a BIACORE® 4000 device (GE Healthcare). Culture supernatant containing variants expressed in FS293-F cells was prepared in ACES buffer, pH 7.4, containing 10 mg/ml BSA and 0.1 mg/ml carboxymethyl dextran (CMD). Each antibody was "captured" on the flow cells until an immobilization level of about 200 RU was reached. Then 25nM human latent myostatin was injected into the flow cells. In the BIACORE® assay, an additional dissociation phase was performed at pH 5.8 immediately after the dissociation phase at pH 7.4. In this additional dissociation phase, the pH-dependent dissociation of the antibody (Ab) and antigen (Ag) from the complexes obtained at pH 7.4 was evaluated. The flow cell surface was regenerated with 25 mM NaOH. Kinetic parameters were determined using BIACORE® 4000 Evaluation software version 2.0 (GE Healthcare). Additional dissociation analysis at pH 5.8 was performed using Scrubber2 (BioLogic software).

Variants with increased affinity at pH 7.4 and/or in vitro inhibitory activity were selected and combined with the pH enhancing mutations identified in Example 11. After combining these mutations, 4 variants (MS1032LO01, 02, 03 and expressed with SG1 or F760 (SEQ ID NOs: 11 and 51) for in vitro and/or in vivo kinetic binding assays with BIACORE®. Both SG1 and F760 are human heavy chain constant regions. SG1 binding affinities to human Fc-gamma R are comparable to the constant region affinity of naturally occurring IgG1, but F760 affinity is eliminated due to Fc modification (referred to herein also as Fc silencing). Amino acid and nucleotide sequences of four variants of antibodies to myostatin are presented in table 2a.

Example 13

Characterization of pH-Dependent Variants of MS 1032

(HEK Blue analysis (BMP1 and spontaneous activation))

All experimental conditions were as described in Examples 3 and 4. As shown in FIG. 10, all variants showed inhibitory activity comparable to MS1032LO00-SG1 against human latent myostatin.

Example 14

Characterization of pH-Dependent Variants of MS 1032 (BIACORE®)

All experimental conditions were as described in Example 7, except that measurements were also made in ACES buffer, pH 5.8, in addition to ACES conditions, pH 7.4. The activity of some antibodies was evaluated only against human latent myostatin. The antibody immobilization level should correspond to 185 RU and 18.5 RU for avidity and affinity analysis, respectively. Kinetic parameters were determined by fitting the data to a 1:1 binding model using BIACORE® T200 Evaluation software version 2.0 (GE Healthcare). Sensorgrams for all anti-human latent myostatin antibodies obtained under the conditions used to evaluate avidity are shown in FIG. 11, a ka, kd, and KD calculated for various conditions are presented in tables 7-10. Under the conditions used to evaluate avidity, all antibodies except MS1032LO00-SG1 showed a faster rate of dissociation at acidic pH than at neutral pH. Under the conditions used to evaluate the affinity, all antibodies except MS1032LO00-SG1 showed little interaction with latent myostatin at acidic pH and therefore kinetic parameters could not be determined.

N/T: not tested

N/T: not tested

N/T: not tested

n.d.: not determined, N/T: not tested

Example 15

Comparison of the concentration of total myostatin in the plasma of mice when using antibodies with the ability to bind to Fc-gamma R, and with eliminated ability to bind to Fc-gamma R

In vivo experiment using C. B-17 SCID mice

The accumulation of endogenous myostatin was assessed in vivo after administration of an antibody to latent myostatin in C.B-17 SCID mice (In Vivos, Singapore). Antibody to latent myostatin (3 mg/ml) was administered as a single dose of 10 ml/kg into the caudal vein. Blood samples were collected at 5 minutes, 7 hours, 1 day, 2 days, 3 days, 7 days, 14 days, 21 days and 28 days after administration. The collected blood was immediately centrifuged at 14,000 rpm at 4° C. for 10 minutes to separate the plasma. The separated plasma was stored at -80° C. or below until analysis. MS1032LO00-SG1 and MS1032LO00-F760 were used as antibodies to latent myostatin.

Plasma measurement of total myostatin by electrochemiluminescence (ECL)

The concentration of total myostatin in the plasma of mice was measured using ECL. Anti-mature myostatin antibody immobilized plates were prepared by applying anti-mature myostatin antibody RK35 (described in WO 2009/058346) to a 96-well MULTI-ARRAY plate (Meso Scale Discovery) and incubating overnight at 4°C. Mature myostatin samples were prepared to obtain a calibration curve and plasma samples diluted 40 times or more. Samples were mixed in an acidic solution (0.2 M glycine-HCl, pH 2.5) to separate by dissociation of mature myostatin from its binding protein (such as a propeptide). The samples were then loaded onto a plate immobilized with anti-mature myostatin antibody and allowed to bind for 1 hour at room temperature, followed by washing. The SULFO-labeled anti-mature myostatin antibody RK22 (described in WO 2009/058346) was then added and the plate was incubated for 1 hour at room temperature followed by washing. Read Buffer T (×4) (Meso Scale Discovery) was immediately added to the plate, and signal detection was performed using a SECTOR Imager 2400 (Meso Scale Discovery). Mature myostatin concentration was calculated from the response to the standard curve using SOFTmax PRO analytical software (Molecular Devices). The time dependence of the plasma concentration of total myostatin after intravenous administration of antibodies to latent myostatin, determined using this method, is shown in Fig. 12.

Effect of Fc-gamma R binding on myostatin accumulation in vivo

After administration of the MS1032LO00-F760 antibody, the plasma concentration of total myostatin accumulated by day 28 was 248-fold higher than the plasma concentration of total myostatin after 5 minutes. In contrast, after administration of MS1032LO00-SG1, the plasma concentration of total myostatin accumulated by day 28 was 37 times the plasma concentration of total myostatin after 5 minutes. Approximately 7-fold difference in plasma concentration of total myostatin at day 28 was found between MS1032LO00-F760 (silent Fc) and MS1032LO00-SG1 as a result of binding to Fc-gamma R. In the case of human soluble IL-6R (hsIL-6R), no significant differences in plasma concentration of hsIL-6R when using antibodies to hsIL-6R F760 (silent Fc) and SG1, as described in WO 2013/125667. Because hsIL-6R is a monomeric antigen, the antibody-hsIL-6R complex contains only one Fc. Thus, the results obtained in WO 2013/125667 suggest that the single Fc antibody-antigen complex does not bind to the Fc-gamma R in vivo to such an extent that the immune complex is taken up by the cells. On the other hand, with a multimeric antigen (such as myostatin), an antibody can form a large immune complex, and the antibody-antigen complex contains more than two Fc. Thus, there may be a significant difference in plasma antigen concentration between F760 and SG1 due to the strong avidity of binding to Fc-gamma R. This result suggests that MST1032 may form a large immune complex with myostatin, which contains more than two antibodies.

Example 16

Comparison of Plasma Concentration of Total Myostatin with pH-Independent Anti-Latent Myostatin Antibody and pH-Dependent Anti-Latent Myostatin Antibody in Mice

In vivo experiment using C.B-17 SCID mice

Endogenous myostatin accumulation was assessed in vivo after administration of anti-myostatin latent antibody to C.B-17 SCID mice (In Vivos, Singapore) according to the method described in Example 15. Anti-myostatin latent antibodies MS1032LO01-SG1 and MS1032LO01-F760 were used.

Plasma total myostatin concentration measurement using ECL

Plasma total myostatin concentration in mice was measured by ECL according to the method described in Example 15. The time course of plasma total myostatin concentration after intravenous administration of anti-latent myostatin antibody, determined using this method, is shown in FIG. 13.

Effect of pH-Dependent Myostatin Binding on Myostatin Accumulation in Vivo

The pH-dependent antibodies to latent myostatin (MS1032LO01-SG1 and MS1032LO01-F760) were tested in vivo and plasma concentrations of total myostatin were compared. Total myostatin concentration data after administration of MS1032LO00-SG1 and MS1032LO00-F760 are shown in Example 15 and also in FIG. 13. MS1032LO00 is a pH independent antibody and MS1032LO01 is a pH dependent antibody. As shown in FIG. 13, total myostatin concentrations after administration of MS1032LO01-F760 decreased compared to MS1032LO00-F760 due to pH dependent binding. In addition, the concentrations of total myostatin after administration of MS1032LO01-SG1 was sharply reduced compared to MS1032LO00-SG1 due to pH-dependent binding and increased penetration into cells as a result of binding to Fc-gamma R. It can be expected that MS1032LO01-SG1 has a very high the ability to increase the clearance of myostatin from plasma as a "Sweeping®-antibody" (antibody-cleaner) (i.e., an antibody that has the ability to actively eliminate soluble antigens from the bloodstream").

Example 17

In vivo efficacy of a pH-dependent antibody to latent myostatin

All experimental designs used were as described in Example 8. As shown in FIG. 14, after intravenous administration to SCID mice at doses of 0.2, 1, and 5 mg/kg, MS1032LO00-SG1 (non-cleaner antibody) and MS1032LO01-SG1 (cleaner antibody) increased skeletal muscle mass in a dose-dependent manner after 2 weeks. MS1032LO01-SG1 antibody significantly increased quadriceps wet mass and grip strength at 1 and 5 mg/kg and significantly increased lean body mass and calf wet mass at 5 mg/kg compared to treated filler (PFR) group. MS1032LO01-SG1 also significantly reduced fat mass at 1 and 5 mg/kg doses relative to the vehicle-treated (PBS) group. MS1032LO00-SG1 at 0.2mg/kg did not show any increase in lean body mass. On the other hand, MS1032LO01-SG1, when administered at a dose of 0.2 mg/kg, dramatically increased lean body mass, quadriceps wet mass, wet mass, and gastrocnemius contraction force. Thus, an antibody with pH-dependent binding characteristics enhanced muscle growth and muscle strength to a greater extent than an antibody with pH-independent binding.

Example 18

Expression and purification of human and mouse latent GDF11

Human GDF11 with a Flag tag at the N-terminus (also referred to herein as Flag-hGDF11, human latent GDF11, hGDF11 or human GDF11, SEQ ID NO: 85) was transiently expressed using FreeStyle293-F cells (Thermo Fisher). Conditioned media expressing Flag-hGDF11 were used to pack a column with anti-Flag M2 affinity resin (Sigma) and eluted with Flag peptide (Sigma). Fractions containing Flag-hGDF11 were collected and then subjected to gel filtration on a Superdex 200 column (GE healthcare) equilibrated with 1×PBS. Fractions containing Flag-hGDF11 were then pooled and stored at -80°C.

Example 19

Characterization of Anti-Latent Myostatin Antibody (ELISA)

Non-sensitized ELISA plates (NUNC-IMMUNO plate, MAXISORP surface, Nalge Nunc International) were coated with 20 μl of 50 nM streptavidin (GenScript) for 2 hours at room temperature. The plates were then washed three times with PBS and blocked overnight with 50 μl of 20% Blocking One (Nacalai Tesque). The next day, each well of the plate was incubated with biotinylated human latent myostatin at 2 nM/well/20 µl or biotinylated human latent GDF11 at 20 nM/well/20 µl for 2 hours. After washing, 20 µl of antibody sample was added to the wells and the plates were incubated for 1 h. The plates were washed and 20 μl of anti-human IgG conjugated to horseradish peroxidase (HRP) (Abeam) diluted in HEPES buffered saline was added, and the plates were incubated for another 1 h. Then the plates were again washed and thereafter, 50 μl of ABTS (KPL) was added to each well, and the plates were incubated for 1 hour. Signal detection was performed at 405 nm in a colorimetric plate reader. The results of the binding experiment are shown in FIG. 15. MST1032-G1m was found to bind to latent myostatin (ie, a non-covalent complex of mature myostatin and propeptides) but not to hGDF11. These results demonstrate that MST1032-G1m specifically binds to myostatin but does not bind to hGDF11.

Example 20

Characterization of Anti-Latent Myostatin Antibody (HEK Blue Assay (BMP1 and Spontaneous Activation))

Reporter gene analysis was used to assess the biological activity of active GDF11 in vitro. HEK-Blue™ TGF-beta cells (Invivogen), which express Smad3/4-binding element (SBE)-indicated SEAP reporter genes, allow for the detection of bioactive GDF11 by monitoring activation of activin type 1 and type 2 receptors. Active GDF11 stimulates production and secretion of SEAP into the cell supernatant. The amount of SEAP secreted is then determined using QUANTIBlue™ (Invivogen).

HEK-Blue™ TGF-beta cells were maintained in DMEM (Gibco) supplemented with 10% fetal bovine serum, 50 µg/ml streptomycin, 50 U/ml penicillin, 100 µg/ml Normocin™, 30 µg/ml blasticidin, 200 µg/ml HygroGold™ and 100 µg/ml Zeocin™. When performing a functional assay, cell medium was changed to assay medium (DMEM supplemented with 0.1% bovine serum albumin, streptomycin, penicillin, and Normocin™) and plated in a 96-well plate. Human GDF11 was incubated with or without recombinant human BMP1 (Calbiochem) and latent myostatin antibody (MST1032-G1m) at 37°C overnight. Sample mixtures were transferred into cells. After a 24 hour incubation, cell supernatants were mixed with QUANTIBlue™ and absorbance at 620 nm was measured in a colorimetric plate reader. As shown in FIG. 16, MST1032-G1m did not interfere with protease-mediated activation or spontaneous activation of human GDF11 and therefore did not inhibit SEAP secretion.

Example 21

Additional optimization of MS1032 variants to increase the "sweeping effect" (cleaner effect)

Since the pH dependent antibody, MS1032LO01-SG1, has been shown to have very high potency, when studied in mice, further optimizations were made to increase pH dependence, increase cell uptake, increase stability, etc. by introducing mutations in the CDR of the antibody. or by changing the frame sections. Over a thousand variants were evaluated using the BIACORE® and/or HEK Blue assays described above and generated MS1032LO06-SG1, MS1032LO07-SG1, MS1032LO10-SG1, MS1032LO11-SG1, MS1032LO12-SG1, MS1032LO18-SG1, MS1032LO19-SG1 , MS1032LO21-SG1, MS1032LO23-SG1, MS1032LO24-SG1, MS1032LO25-SG1 and MS1032LO26-SG1. The amino acid and nucleotide sequences of these generated antibodies are shown in Table 11.

The binding affinity of MS1032 variants to latent human, cynomolgus (cyno) monkeys and mice myostatin was assessed at 37° C. at pH 7.4 and pH 5.8 using a BIACORE® T200 device (GE Healthcare) to evaluate the effect of pH on antigen binding. . ProA/G (Pierce) was immobilized on all flow cells of the CM4 chip using an amine coupling kit (GE Healthcare). All antibodies and analytes were prepared in ACES buffer, pH 7.4 or pH 5.8, containing 20 mM ACES, 150 mM NaCl, 1.2 mM CaCl ₂ , 0.05% Tween 20, 0.005% NaN ₃ . Each antibody was "captured" on the surface of the sensor chip using ProA/G. Antibody immobilization levels typically corresponded to 130-240 resonance units (RU). Human, cyno, or mouse latent myostatin was injected at concentrations ranging from 3.125 to 50 nM, which were obtained by two-fold serial dilutions and then allowed to dissociate. The sensor surface was regenerated after each cycle with 10 mM glycine-HCl, pH 1.5. Binding affinity was determined by processing and fitting the data to a 1:1 binding model using BIACORE® T200 Evaluation software version 2.0 (GE Healthcare).

Binding affinity (KD) data for MS1032 variants with latent human, cyno, and mouse myostatin at pH 7.4 and pH 5.8 are shown in Table 12. For all variants, the ratio of KD values ((KD at pH 5.8)/( KD at pH 7.4)) of more than 5, indicating pH-dependent binding to latent myostatin.

Example 22

Evaluation of the neutralizing activity of the variants obtained as a result of additional optimization using the HEK Blue analysis

Neutralizing activity of MS1032LO06-SG1, MS1032LO07-SG1, MS1032LO10-SG1, MS1032LO11-SG1, MS1032LO12-SG1, MS1032LO18-SG1, MS1032LO19-SG1, MS1032LO21-SG1, MS1032LO23-SG1 and MS1032LO25-SG1, MS1032LO23-SG1 and MS1032LO25-SG1 latent human ostatin using the method described in Example 3. As shown in FIG. 17, all variants showed activity comparable to that of MS1032LO01-SG1.

Example 23

Comparison of total myostatin plasma concentration in mice using a pH-independent antibody to latent myostatin and various pH-dependent antibodies to latent myostatin

In vivo experiment using C.B-17 SCID mice

The accumulation of endogenous myostatin was assessed in vivo after administration of an antibody to latent myostatin in C.B-17 SCID mice (In Vivos, Singapore). Antibody to latent myostatin (3 mg/ml) was administered as a single dose of 10 ml/kg into the caudal vein. Blood samples were collected at 5 minutes, 7 hours, 1 day, 2 days, 3 days, 7 days, 14 days, 21 days and 28 days after administration. The collected blood was immediately centrifuged at 14,000 rpm at 4° C. for 10 minutes to separate the plasma. The separated plasma was stored at -80° C. or below until analysis. MS1032LO00-SG1, MS1032LO01-SG1, MS1032LO06-SG1, MS1032LO11-SG1, MS1032LO18-SG1, MS1032LO19-SG1, MS1032LO21-SG1 and MS1032LO25-SG1 were used as antibodies to latent myostatin.

Plasma measurement of total myostatin by electrochemiluminescence (ECL)

The concentration of total myostatin in the plasma of mice was measured using ECL. Anti-mature myostatin antibody immobilized plates were prepared by applying biotinylated anti-mature myostatin antibody RK35 (described in WO 2009/058346) to a 96-well streptavidin-coated MULTI-ARRAY plate (Meso Scale Discovery) and incubating in blocking buffer for 2 hours at room temperature. Mature myostatin samples were prepared to obtain a calibration curve and plasma samples diluted 40 times or more. Samples were mixed in an acidic solution (0.2 M glycine-HCl, pH 2.5) to separate by dissociation of mature myostatin from its binding protein (such as a propeptide). The samples were then loaded onto a plate immobilized with anti-mature myostatin antibody and allowed to bind for 1 hour at room temperature, followed by washing. Then SULFO labeled anti-mature myostatin antibody RK22 (described in WO 2009/058346) was added and the plate was incubated for 1 hour at room temperature followed by washing. Read Buffer T (×4) (Meso Scale Discovery) was immediately added to the plate and signal was detected using a SECTOR Imager 2400 (Meso Scale Discovery). Mature myostatin concentration was calculated from the response to the standard curve using SOFTmax PRO analytical software (Molecular Devices). The time dependence of the plasma concentration of total myostatin after intravenous administration of antibodies to latent myostatin, determined using this method, is shown in Fig. 18.

Effect of pH-Dependent Binding to Myostatin on Myostatin Accumulation in Mice in an In Vivo Experiment

The effect of pH-dependence on myostatin accumulation in mice was compared using a pH independent latent myostatin antibody (MS1032LO00-SG1) and various pH dependent latent myostatin antibodies (MS 1032LO01-SGI, MS 1032LO06-SG1, MS 1032LO11-SGI, MS1032LO18 -SG1, MS1032LO19-SG1, MS1032LO21-SG1 and MS1032LO25-SG1). The introduction of pH dependence significantly accelerated the clearance of myostatin from the plasma of SOD mice. As shown in FIG. 18, pH-dependent antibodies to latent myostatin (MS1032LO01-SG1, MS1032LO06-SG1, MS1032LO11-SG1, MS1032LO18-SG1, MS1032LO19-SG1, MS1032LO21-SG1, and MS1032LO25-SG1) could reduce myostatin accumulation compared to a pH-independent antibody to latent myostatin (MS1032LO00-SG1) on day 28.

Example 24

Comparison of total myostatin concentration in plasma of cynomolgus monkeys using a pH-independent antibody to latent myostatin and pH-dependent antibodies to latent myostatin having an engineered Fc region

In vivo experience using cynomolgus monkeys

The accumulation of endogenous myostatin was assessed in vivo after administration of antibodies to latent myostatin to 2-4-year-old Masaca fascicularis (cynomolgus cynomolgus monkey) from Cambodia (Shin Nippon Biomedical Laboratories Ltd., Japan). A dose of 30 mg/kg was injected into the cephalic vein of the forearm using a disposable syringe, an extension tube, an internal needle and an infusion pump. The dosing rate was 30 minutes per animal. Blood samples were collected before the start of dosing and either 5 min, 7 h and 1, 2, 3, 7, 14, 21, 28, 35, 42, 49 and 56 days after the end of dosing, or 5 min and 2, 4 and 7 hours and 1, 2, 3, 7, 14, 21, 28, 35, 42, 49 and 56 days after the end of dosing. Blood was taken from the femoral vein using a syringe containing sodium heparin. The blood was immediately cooled on ice and plasma was obtained by centrifugation at 4°C, 1700×g for 10 min. Plasma samples were stored in a deep freezer (acceptable range: -70° C. or below) until analysis. В качестве антител к латентному миостатину применяли MS1032LO00-SG1, MS1032LO06-SG1012, MS1032LO06-SG1016, MS1032LO06-SG1029, MS1032LO06-SG1031, MS1032LO06-SG1033 и MS1032LO06-SG1034 (в контексте настоящего описания SG1012, SG1016, SG1029, SG1031, SG1033 и SG1034 denote heavy chain constant regions constructed from SG1 according to the method described below).

Anti-latent myostatin antibodies MS 1032LO19-SG1079, MS1032LO19-SG1071, MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081 and MS1032LO19-SG1074, MS1032LO19-SG1081 and SG1077 (as used herein, SG1079, SG1071, SG1080, SG1074, SG1081, and SG1077 denote heavy chain constant regions constructed from SG1 according to the method described below). Blood samples were collected before dosing and 5 minutes and 2, 4 and 7 hours, and 1, 2, 3, 7, 14 days after the end of dosing. The blood was processed according to the procedure described above. Plasma samples were stored in a deep freezer (acceptable range: -70° C. or below) until analysis.

Plasma measurement of total myostatin by electrochemiluminescence (ECL)

Plasma total myostatin concentration in monkeys was measured by ECL according to the method described in Example 23. The time course of plasma total myostatin concentration after intravenous administration of anti-latent myostatin antibody, determined using this method, is shown in FIG. 19.

Measurement of monkey plasma ADA by electrochemiluminescence (ECL)

The biotinylated drug was used to sensitize a 96-well MULTI-ARRAY streptavidin plate (Meso Scale Discovery) and incubated in LowCross® buffer (Candor) for 2 hours at room temperature. Monkey plasma samples were diluted 20-fold in LowCross® buffer prior to plate loading. Samples were incubated overnight at 4°C. The next day, the plate was washed three times with wash buffer before adding a SULFO-labeled anti-monkey IgG secondary antibody (Thermo Fisher Scientific). After incubation for 1 hour at room temperature, the plate was washed three times with wash buffer. Read Buffer T (x4) (Meso Scale Discovery) was immediately added to the plate, and signal detection was performed with a SECTOR Imager 2400 (Meso Scale Discovery).

Effect of pH Dependence and Fc Engineering on Myostatin Accumulation in Monkeys in Vivo

Administration of pH-independent antibody (MS1032LO00-SG1) to cynomolgus monkeys resulted in at least a 60-fold increase in myostatin concentrations from baseline on day 28. On day 28, pH-dependent latent myostatin antibodies MS1032LO06-SG1012 and MS1032LO06-SG1033 provided and 8-fold increase from baseline, respectively. The strong swipping effect is mainly associated with an increase in affinity for Fc-gamma RIIb of cynomolgus monkeys. On day 28, pH-dependent latent myostatin antibodies MS1032LO06-SG1029, MS1032LO06-SG1031 and MS1032LO06-SG1034 were able to eliminate the antigen to below baseline levels. The reason for the strong swipping effect in the case of MS1032LO06-SG1029, MS1032LO06-SG1031 and MS1032LO06-SG1034 is an increase in non-specific uptake by the cell due to an increase in the positively charged antibody cluster and an increase in Fc-gamma R mediated cellular uptake due to increased binding to Fc-gamma R .

The introduction of pH-dependent antibodies to latent myostatin MS1032LO19-SG1079, MS1032LO19-SG1071, MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081 and MS1032LO19-SG1077 reduced the concentration of myostatin to a level below the detection limit (<0.25 ng/mL), starting from from day 1 in cynomolgus monkeys. On day 14, myostatin concentrations exceeded the detection limit with MS1032LO19-SG1079 and MS1032LO19-SG1071, although myostatin levels remained below the detection limit with MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081, and MS1032LO19-SG1077. The weaker suppression found for MS1032LO19-SG1079 and MS1032LO19-SG1071 may be the result of various mutations leading to a change in pI.

The results suggest that a strong elimination of myostatin from plasma can be achieved by mutations that increase binding to Fc-gamma RIIb or combinations of mutations that increase the positive charge of the antibody and increase binding to Fc-gamma RIIb. It can be expected that a strong elimination of myostatin can be achieved in humans by combining mutations that increase the positive charge of the antibody and increase binding to Fc-gamma R.

Example 25

In vivo efficacy of additionally optimized variants in mice

As described in Example 8, in vivo efficacy was evaluated in Scid mice using MS1032LO06-SG1, MS1032LO11-SG1, MS1032LO18-SG1, MS1032LO19-SG1 and MS1032LO25-SG1. Three independent experiments were carried out and MS1032LO01 was used as a control.

The results of these experiments are shown in Fig. 20A-20I. All antibodies (MS1032LO06-SG1, MS1032LO11-SG1, MS1032LO18-SG1, MS1032LO19-SG1 and MS1032LO25-SG1) were found to increase lean body mass (LBM) as well as appendicular muscle contraction force. All antibodies also showed a decrease in body fat mass in a dose-dependent manner.

Example 26

Epitope mapping of antibodies to latent myostatin

Additional antibodies to human latent myostatin were designed to map their respective epitopes to which they bind. Two NZW rabbits were immunized and antibodies were obtained according to the method described in Example 2. The 1760 antibody-producing B cell lines identified were further screened for their ability to block BMP1 mediated activation of human latent myostatin. In general, the method was as follows: B cell supernatant containing secreted antibodies was incubated with human latent myostatin in the presence of recombinant human BMP1 (R&D Systems) at 37°C overnight. Then 50 μl of the reaction mixture was transferred into a 96-well MULTI-ARRAY plate from Meso Scale Discovery sensitized with an antibody to mature myostatin RK35 (described in WO 2009/058346). After incubation for 1 hour at room temperature with shaking, the plates were incubated with biotinylated anti-mature myostatin antibody RK22 (described in WO 2009/058346) and then with SULFO-labeled streptavidin. Then, Read Buffer T (×4) (Meso Scale Discovery) was added to the plate, and signal detection was performed using a SECTOR Imager 2400 (Meso Scale Discovery). For subsequent analysis, 94 lines were selected, characterized by different levels of neutralizing activity (MST1495-MST1588). The variable regions of these selected lines were cloned according to the method described in Example 2, except that an expression vector with the heavy chain constant region sequence F1332m (SEQ ID NO: 193) was used.

Inhibitory activity against human latent myostatin activation was further evaluated for a panel of 7 anti-latent myostatin antibodies (MST1032, MST1504, MST1538, MST1551, MST1558, MST1572, and MST1573; the identifying amino acid sequence numbers of these antibodies are shown in Table 13). As shown in FIG. 21, all antibodies were able to inhibit BMP1 mediated activation of human latent myostatin in a dose dependent manner.

Four antibodies that showed promise by Western blot analysis were selected for epitope mapping. Fragments of the N-terminal propeptide coding region of human latent myostatin were cloned into the pGEX4.1 vector so as to obtain GST labeled propeptide fragments each of 100 amino acids with 80 overlapping amino acids (FIG. 22A). Protein expression was induced in BL21-transformed competent cells using the Overnight Express Autoinduction System (Merck Millipore) and the protein was extracted with BugBuster Protein Extraction Reagent (Novagen). Expression of the desired 37 kDa protein fragments was confirmed by Western blot analysis as described in Example 6 (anti-GST antibody (Abeam)) (FIG. 22B). When examining antibodies to human latent myostatin indicated in FIG. 22B, while all 4 antibodies could inhibit latent myostatin activation, they recognized different epitopes on human latent myostatin. The MST1032 antibody could detect the first five fragments, no bands were detected after deletion of amino acids 81-100 (SEQ ID NO: 78). Both antibodies MST1538 and MST1572 bound only to the first three fragments, no bands were detected in the absence of amino acids 41-60 of SEQ ID NO: 78. The MST1573 antibody strongly bound only to the first two fragments, which suggests that its epitope lies in amino acids 21-40 SEQ ID NO: 78 (FIG. 22D).

Example 27

Creation of new Fc variants with increased activity against Fc-gamma RIIb

This example illustrates the construction of Fc to increase myostatin clearance.

WO 2013/125667 demonstrates that the clearance of a soluble antigen can be increased by administering antigen-binding molecules containing an Fc domain with increased affinity for Fc-gamma RIIb. In addition, Fc variants that have been found to have increased ability to bind to human Fc-gamma RIIb are illustrated in WO 2012/115241 and WO 2014/030728. In addition, it has been demonstrated that these Fc variants can have a selectively increased ability to bind to human Fc-gamma RIIb and a reduced ability to bind to other active Fc-gamma R. This selective increase in binding to Fc-gamma RIIb can have a beneficial effect not only on clearance of soluble antigen, but also to reduce the risk of unwanted effector functions and immune response.

When developing an antibody as a drug, efficacy, pharmacokinetics, and safety should be studied in non-human animals for which the drug has pharmacological activity. If it is only active in humans, alternative approaches should be used, such as using a surrogate antibody (Int. J. Tox. 28, 2009, pp. 230-253). However, it is difficult to predict the effects of the interaction between the Fc region and the Fc-gamma R in humans using a surrogate antibody, because the Fc-gamma R expression and/or function patterns in non-human animals are not always the same as in humans. Preferably, the Fc regions of antibodies used as drugs have cross-reactivity with regions of animals other than humans, especially cynomolgus monkeys, which have expression patterns and functions of Fc-gamma R close to those of humans, so that results obtained in animals other than a person can be extrapolated to a person.

Therefore, when creating the present invention created Fc-variants with cross-reactivity between Fc-gamma R human and cyno.

Measurement of the affinity of existing Fc variants with increased activity against Fc-gamma RIIb for human and simian Fc-gamma R

Heavy chain gene of a variant with increased activity against human Fc-gamma RIIb (in the present description, the term "with increased activity against Fc-gamma RIIb" means "with increased Fc-gamma RIIb-binding activity"), described in WO 2012/115241 , was created by replacing Pro at position 238 according to EU numbering with Asp in the MS1032LO06-SG1 heavy chain, which was designated M103205H795-SG1 (VH, SEQ ID NO: 86; CH, SEQ ID NO: 9). The resulting heavy chain was designated M103205H795-MY009 (VH, SEQ ID NO: 86; CH, SEQ ID NO: 252). M103202L889-SK1 (VL, SEQ ID NO: 96; CL, SEQ ID NO: 10) was used as the light chain of the antibody. The recombinant antibody was expressed according to the method described in Example 34.

The extracellular domains of Fc-gamma R were obtained as follows. Synthesis of human Fc-gamma R extracellular domain genes was performed using methods known to those skilled in the art based on information registered with the NCBI. In particular, the basis for creating Fc-gamma RIIa was the NCBI sequence with access code No. NM_001136219.1, Fc-gamma RIIb - NM_004001.3, Fc-gamma RIIIa - NM_001127593.1, respectively. The Fc-gamma RIIa and Fc-gamma RIIIa allotypes were derived from the Fc-gamma RIIa (J. Exp. Med. 172, 1990, pp. 19-25) and Fc-gamma RIIIa (J. Clin. Invest. 100) polymorphism data. (5), 1997, pp. 1059-1070) respectively. Synthesis of the cynomolgus Fc-gamma R extracellular domain gene was accomplished by cloning the cDNA of each cynomolgus Fc-gamma R using methods known to those skilled in the art. The amino acid sequences of the engineered extracellular domains of Fc-gamma R are shown in the sequence listing: (SEQ ID NO: 210 for human Fc-gamma RIIaR, SEQ ID NO: 211 for human Fc-gamma RIIaH, SEQ ID NO: 214 for human Fc-gamma RIIb , SEQ ID NO: 217 for human Fc-gamma RIIIaF, SEQ ID NO: 218 for human Fc-gamma RIIIaV, SEQ ID NO: 220 for cyno Fc-gamma RIIa1, SEQ ID NO: 221 for cyno Fc-gamma RIIa2, SEQ ID NO: 222 for cyno Fc-gamma RIIa3, SEQ ID NO: 223 for cyno Fc-gamma RIIb, SEQ ID NO: 224 for cyno Fc-gamma RIIIaS). A His tag was then added to their C-terminus and each resulting gene was inserted into an expression vector designed for expression in mammalian cells. The expression vector was introduced into FreeStyle293 human embryonic kidney cells (Invitrogen) to express the desired protein. After cultivation, the resulting culture supernatant was filtered and purified mainly by the following four purification steps. Cation exchange chromatography using SP Sepharose FF was performed as the first step, anti-His tag affinity chromatography (HisTrap HP) was performed as the second step, gel filtration on columns (Superdex200) was used as the third step, and sterilization by filtration was used as the fourth step. . The absorbance at 280 nm of the purified proteins was measured with a spectrophotometer and the concentration of the purified protein was determined using the extinction coefficient calculated by the PACE method (Protein Science 4, 1995, pp. 2411-2423).

Kinetic analysis of interactions between these antibodies and Fc-gamma R was performed using BIACORE® T200 or BIACORE® 4000 (GE Healthcare). HBS-EP+ (GE Healthcare) was used as running buffer and the measurement temperature was set to 25°C. The chip was prepared by immobilizing protein A, protein A/G or mouse kappa light chain to human IgG (BD Biosciences) on an S series CM4 or CM5 sensor chip (GE Healthcare) using the amine coupling method. The antibody of interest was "captured" on the surface of the chip for interactions with each Fc-gamma R that was diluted in running buffer and binding to the antibody was measured. After the measurement, the antibody immobilized on the chip was washed off, allowing it to react with 10 mM glycine-HCl, pH 1.5, and 25 mM NaOH, resulting in chip regeneration and reuse. The obtained sensograms, which are the results of the measurement, were analyzed using a 1:1 Langmuir binding model using the BIACORE® Evaluation software to calculate the binding rate constant ka (l/mol/s) and the dissociation reaction rate constant kd (1/s), and on Based on these values, the dissociation constant KD (mol/l) was calculated. Since the binding of MS1032LO06-MY009 to human Fc-gamma RIIaH, human Fc-gamma RIIIaV and cynoFc-gamma RIIIaS was weak, kinetic parameters such as KD could not be calculated using the above analytical method. The KD values describing these interactions were calculated using the 1:1 binding model described below in the BIACORE® T100 Handbook BR1006-48 Edition AE software.

The behavior of interacting molecules based on the BIACORE® 1:1 binding model can be described by equation 1: Req=C×Rmax/(KD+C)+RI where, Req denotes the functional dependence of stationary binding levels on analyte concentration, C denotes concentration, RI is the total refractive index contribution for the sample, and Rmax is the analyte binding capacity of the surface. If we transform the above equation, then we can express KD in the form of equation 2: KD=C×Rmax/(Req-RI)-C. KD can be calculated by substituting the appropriate values for the parameters Rmax, RI and C in Equation 1 or Equation 2. For the measurement conditions considered, the values RI=0, C=2 µmol/l can be used. In addition, the Rmax value obtained by a global fit of the sensogram measured by analyzing the interaction of each Fc-gamma R with IgG1 using a 1:1 Langmuir binding model was divided by the amount of SG1 entrapped, the result was multiplied by the amount of MY009 entrapped, and the resulting value was used as Rmax. This calculation is based on the hypothesis that the limit on the amount of each Fc-gamma R that can be associated with SG1 remains unchanged for all variants obtained by introducing mutations in SG1, and that the value of Rmax at the time of measurement is proportional to the amount of antibody bound with the chip at the time of the measurement. Req was defined as the number of bindings of each Fc-gamma R to each variant on the sensor chip detected at the time of the measurement.

Table 14 shows the results of a kinetic analysis of the binding of SG1 and MY009 to human and cynoFc-gamma R. KD values in cells in bold were calculated using Equation 2 because their affinity was too low to be accurately determined by kinetic analysis. The fold of KD values was calculated by dividing the KD value for SG1 by the KD value for the variant for each Fc-gamma R.

As shown in Table 14, the MY009 variant with increased activity against human Fc-gamma RIIb did not show increased binding to cynoFc-gamma RIIb, but increased binding to human Fc-gamma RIIb. Its affinity for cynoFc-gamma RIIb even turned out to be reduced by 0.4 times compared to SG1, which indicates that the MY009 variant with increased activity against human Fc-gamma RIIb does not cross-react to cynoFc-gamma R.

Development of a novel Fc variant with increased binding to both human and cynoFc-gamma RIIb

Ideally, the new Fc variant should have increased selective binding to both human and cynoFc-gamma RIIb and reduced binding to other active Fc-gamma R. However, since the putative residues responsible for Fc-binding in cynoFc-gamma RIIb are fully consistent with the residues in any cynoFc-gamma RIIa allotype (FIG. 23), it is theoretically impossible to selectively increase binding to cynoFc-gamma RIIb relative to cynoFc-gamma RIIa binding. Thus, the novel Fc variant should have selectively increased binding to human Fc-gamma RIIb, cynoFc-gamma RIIb and cynoFc-gamma RIIa.

To generate a new Fc variant characterized by selectively increased binding to both human and cyno-Fc-gamma RIIb, the sweep mutagenesis (brute force amino acid substitution) assay described in WO 2012/11524 was used. In a sweep mutagenesis assay, sweep mutations were introduced at all positions in the Fc-gamma R binding region of IgG1 isotype antibodies, and a complete binding analysis was performed on each Fc-gamma R according to the procedures below.

The variable region of an anti-glypican 3 antibody containing the CDR GpH 7, which is an antibody to glypican 3 with enhanced plasma kinetics, as described in WO 2009/041062, was used as the variable region of the heavy chain of the antibody (GpH 7: SEQ ID NO: 225 ). Similarly, as the light chain of the antibody, GpL16-k0 (SEQ ID NO: 226), the plasma-enhanced anti-glypican 3 antibody described in WO 2009/041062 was used. In addition, the sequence of the B3 antibody (SEQ ID NO: 228) in which the K439E mutation was introduced into G1d (SEQ ID NO: 227) obtained by deleting the C-terminal Gly and Lys of IgGl was used as the H chain constant region. The specified heavy chain, created by the fusion of GpH 7 and B3, was designated as GpH 7-B3 (VH, SEQ ID NO: 225; CH, SEQ ID NO: 228).

In GpH, 7-B3 amino acid residues that are expected to be involved in binding to Fc-gamma R and their surrounding amino acid residues (positions 234-239, 265-271, 295, 296, 298, 300 and 324-337 according to EU numbering) were replaced by 18 amino acid residues, respectively, excluding the original amino acid residue and Cys. These Fc variants are referred to as B3 variants. B3 variants were expressed and the binding of protein A-purified antibodies to each of the Fc-gamma R (Fc-gamma RIIa type H, Fc-gamma RIIa type R, Fc-gamma RIIb and Fc-gamma RIIIaF) was comprehensively assessed as follows. Analysis of the interaction between each modified antibody and Fc-gamma receptor, obtained according to the method described above, was carried out using BIACOR® T100 (GE Healthcare), BIACORE® T200 (GE Healthcare), BIACORE® A100 or BIACORE® 4000. running buffer used HBS-EP+ (GE Healthcare) and the measurement temperature was set to 25°C. The chip was prepared by immobilizing Protein A (Thermo Scientific), Protein A/G (Thermo Scientific), or Protein L (ACTIGEN or BioVision) on an S-series CM4 or CM5 sensor chip (GE Healthcare) using the amine coupling method. . After capturing the antibodies of interest on these sensor chips, the Fc-gamma receptor diluted in running buffer was allowed to react and the amount of antibody-bound receptors measured. However, since the number of bound Fc-gamma receptors depended on the number of "captured" antibodies, the number of bound Fc-gamma receptors was divided by the number of each "captured" antibody to obtain corrected values, and these values were compared. In addition, the antibodies immobilized on the chips were washed with 10 mM glycine-HCl, pH 1.5, resulting in chip regeneration and reuse. The level of Fc-gamma R binding to each B3 variant was divided by the level of Fc-gamma R binding of the parent B3 antibody (an antibody having the sequence of naturally occurring human IgG1 at positions 234-239, 265-271, 295, 296, 298, 300, and 324-337 according to the EU numbering). The value obtained by multiplying this level by 100 was used as an indication of the relative Fc-gamma R-binding activity of each variant.

Table 15 shows the binding profile of prospective substitutions selected based on the following criteria: more than 40% more binding to human Fc-gamma RIIb, more than 100% less binding to human Fc-gamma RIIaR and Fc-gamma RIIaH, less than than 10% binding to human Fc-gamma RIIIaF compared to parental B3 antibody.

The next step was to assess the cross-reactivity of these substitutions against cynoFc-gamma R. 16 of these mutations were introduced into M103205H795-SG1. Variants were expressed using M103202L889-SK1 as light chain and analyzed for their affinity for cynoFc-gamma RIIa1, IIa2, IIa3, IIb, IIIaS.

Table 16 shows the binding profile of these 16 variants to cynoFc-gamma R. The level of Fc-gamma R binding was obtained by dividing the amount of Fc-gamma R bound by the amount of each antibody captured. This value was standardized taking the value obtained for SG1 as 100.

Of the 16 Fc variants selected, only MY001 retained 85% binding to cynoFc-gamma RIIb relative to SG1 binding. In addition, MY001 showed reduced binding to cynoFc-gamma RIIIaS, while its binding to cynoFc-gamma RIIa1, IIa2 and IIa3 was preserved. As a result, the G236N mutation was found to be a unique substitution that provides a similar binding profile to both human and cynoFc-gamma R.

Although the G236N substitution is cross-reactive with both human and cynoFc-gamma R, when introduced, the affinity for Fc-gamma RIIb is lower compared to SG1 (75% for human Fc-gamma RIIb and 85% for cynoFc-gamma RIIb, respectively). Additional substitutions were evaluated to increase affinity for Fc-gamma RIIb. Specifically, based on the result of the sweep mutagenesis assay (Table 17), substitutions were selected that exhibit increased binding to human Fc-gamma RIIb, reduced binding to human Fc-gamma RIIIa, and increased selectivity for human Fc-gamma RIIb compared to with human Fc-gamma RIIa.

Among these substitutions, L234W and L234Y were selected to improve the selectivity for human Fc-gamma RIIb over human Fc-gamma RIIa. In addition, G236A, G236S, A330R, A330K and P331E were selected to reduce binding to human Fc-gamma RIIIa. All other substitutions were selected to increase binding to human Fc-gamma RIIb. The cross-reactivity of the selected substitutions to cynoFc-gamma R was evaluated. In addition, the three substitutions P396M, V264I and E233D were also evaluated. Substitutions were introduced into M103205H795-SG1 and variants were expressed using M103202L889-SK1 as the light chain.

Table 18 shows the results of the kinetic analysis of selected substitutions, including G236N, in relation to both human and cynoFc-gamma R. In particular, the substitutions were introduced in M103205H795-SG1. Variants were expressed using M103202L889-SK1 as light chain and analyzed for their affinity for cynoFc-gamma RIIa1, IIa2, IIa3, IIb, IIIaS.

The KD values in bold cells were calculated using Equation 2 because their affinity was too low to be accurately determined by kinetic analysis. The fold of KD values was calculated by dividing the KD value for SG1 by the KD value for the variant for each Fc-gamma R.

For G236N, a binding affinity comparable to SG1 was found for cynoFc-gamma RIIa1, cynoFc-gamma RIIa2, cynoFc-gamma RIIa3, and cynoFc-gamma RIIb. Although the affinity for human Fc-gamma RIIb decreased to 0.6, the affinity for human Fc-gamma RIIaH and the affinity for human Fc-gamma RIIaR decreased to 0.2 and 0.1, respectively, indicating that this substitution has high selectivity for human Fc-gamma RIIb compared to human Fc-gamma RIIa. In addition, the affinities for cynoFc-gamma RIIIaS and human Fc-gamma RIIIaV were 0.03 and 0.04, respectively, which is preferable for eliminating ADCC activity. Based on this result, G236N has almost perfect cross-reactivity with human and cynoFc-gamma R, although affinity for both human and cynoFc-gamma RIIb may be increased when administered.

Among the additional substitutions studied, S298L, G236A, P331E, E233D, K334Y, K334M, L235W, S239V, K334I, L234W, K328T, Q295L, K334V, K326T, P396M, I332D, H268E, P271G, S267A and H268D resulted in increased binding to human and H268D , and with cynoFc-gamma RIIb. In particular, H268D showed the highest potency (7-fold for human Fc-gamma RIIb and 5.3-fold for cynoFc-gamma RIIb). Regarding the effect of reducing binding to Fc-gamma RIIIa, less than 0.5-fold affinity was found for G236S, A330K and G236D compared to SG1.

Combinations of the substitutions listed in Table 18 were evaluated to increase the affinity of MY001 for both human and cynoFc-gamma RIIb. In particular, substitutions were introduced in M103205H795-SG1. Variants were expressed using M103202L889-SK1 as light chain and analyzed for their affinity. Table 19 shows the results of a kinetic analysis of binding to human and cynoFc-gamma R. KD values in bold cells were calculated using Equation 2 because their affinity was too low to be accurately determined by kinetic analysis. The fold of KD values was calculated by dividing the KD value for SG1 by the KD value for the variant for each Fc-gamma R.

All variants suppressed affinity for human Fc-gamma RIIIaV by less than 0.26-fold and for cynoFc-gamma RIIIaS by less than 0.42-fold compared to SG1. In addition, for all variants except MY047, MY051 and MY141, increased binding to both human and cynoFc-gamma RIIb was successfully demonstrated compared to MY001, their binding to human Fc-gamma RIIa was maintained at less than 2-fold by compared to SG1. Among them, MY201, MY210, MY206, MY144, MY103, MY212, MY105, MY205, MY109, MY107, MY209, MY101, MY518, MY198 and MY197 were characterized by increased binding to both human and cynoFc-gamma RIIb, exceeding more than 4 fold binding of SG1. In addition, MY205, MY209, MY198 and MY197 showed over 7-fold increased binding to both human and cynoFc-gamma RIIb.

WO 2014030728 illustrates that substitutions at position 396 in the CH3 domain increased affinity for human Fc-gamma RIIb. A complete enumeration of amino acid substitutions was introduced at position 396 M103205H795-MY052, which contains the G236N/H268E/P396M substitutions. The resulting variants were expressed using M103202L889-SK1 as light chain and their affinity for human and cynoFc-gamma R was evaluated (Table 20). The fold of KD values was calculated by dividing the KD value for SG1 by the KD value for the variant for each Fc-gamma R.

Among the substitutions studied, P396I, P396K, and P396L maintained more than 5-fold binding to human Fc-gamma RIIb compared to SG1, and only substitution P396L resulted in increased affinity for human Fc-gamma RIIb compared to MY052. Regarding binding to cynoFc-gamma RIIb, the parental MY052 showed the highest affinity, 3.9 times higher than that of SG1.

Because G236N has ideal cross-reactivity with human and cynoFc-gamma R, other substitutions at position 236 that were not explored in the previous examples were studied. In particular, Asn in M103205H795-MY201 was replaced with Met, His, Val, Gln, Leu, Thr, and Ile. The resulting variants were expressed using M103202L889-SK1 as light + chain and their affinity for human and cynoFc-gamma R was evaluated (Table 21). The KD values in bold cells were calculated using Equation 2 because their affinity was too low to be accurately determined by kinetic analysis. The fold of KD values was calculated by dividing the KD value for SG1 by the KD value for the variant for each Fc-gamma R.

Although all variants had reduced affinity for both human and cynoFc-gamma RIIb compared to the parent molecule MY201, MY265, which was generated by replacing Asn with Thr at position 236 of MY201, retained a 2.1-fold increased binding to human Fc- gamma RIIb and 3.1-fold binding to cynoFc-gamma RIIb, respectively, compared to SG1. The MY265 variant also retained reduced binding to both human and cynoFc-gamma RIIIa. Although the affinity for human Fc-gamma RIIaH and Fc-gamma RIIaR increased more than 2.5-fold compared to SGI, G236T was the second preferred substitution at position 236.

To improve the selectivity and affinity of MY265 for the human Fc-gamma RIIb, a complete enumeration of amino acid substitutions at positions 231 and 232 was carried out. In particular, the amino acid at positions 231 and 232 of M103205H795-MY265 was replaced with one of 18 amino acid residues, excluding the original amino acid and Cys. The resulting variants were expressed using M103202L889-SK1 as light chain and their affinity for human and cynoFc-gamma R was evaluated (Table 22). The fold of KD values was calculated by dividing the KD value for SG1 by the KD value for the variant for each Fc-gamma R.

Regarding affinity for Fc-gamma RIIb, the addition of A231T, A231M, A231V, A231G, A231F, A231I, A231L, A231, A231W, P232V, P232Y, P232F, P232M, P232I, P232W, P232L increased binding to both human and cynoFc- RIIb compared to the parent molecule MY265. Moreover, the addition of A231T and A231G reduced binding to human Fc-gamma RIIaR compared to MY265.

Combinations of substitutions that were not tested in the previous examples were studied. Tested and presumably promising substitutions were introduced into M103205H795-SG1 in combination. The resulting variants were expressed using M103202L889-SK1 as light chain and their affinity for human and cynoFc-gamma R was evaluated. , because their affinity was too low to accurately determine it using kinetic analysis. The fold of KD values was calculated by dividing the KD value for SG1 by the KD value for the variant for each Fc-gamma R. For comparison, the values for MY209 selected as promising variants from the results presented in Table 19 are again shown.

Among the variants tested, only MY213 showed a higher binding affinity for both human and cynoFc-gamma RIIb compared to MY209. However, the affinity of MY213 for human Fc-gamma RIIaH and human Fc-gamma RIIaR was higher than for MY209.

Example 28

Evaluation of myostatin clearance using Fc variants with increased activity against Fc-gamma RIIb in mice transgenic for all human Fc-gamma R

The effect on myostatin clearance of an Fc variant with enhanced activity against Fc-gamma RIIb, constructed according to the method described in Example 27, was evaluated in mice in which all murine Fc-gamma R's were deleted by deletion and human Fc-gamma R's encoded in as transgenes, were inserted into the mouse genome (Proc. Natl. Acad. Sci., 109, 2012, p. 6181). In these mice, all murine Fc-gamma R is replaced by human receptors, so the effect of increased affinity for human Fc-gamma RIIb on soluble antigen clearance can be assessed in these mice. In addition, pI-increasing substitutions were evaluated in combination with Fc variants with increased activity against Fc-gamma RIIb, which were designed according to the method described in Example 27.

Obtaining and profiling test cases

The 8 antibodies tested and their binding profiles are summarized in Table 24. The heavy chain, MS103205H795-PK2, was generated by introducing substitutions that increase pI (S400R/D413K) in MS103205H795-MY101. MS103205H795-MY351, MS103205H795-MY344, MS103205H795-MY335 were obtained by introducing other pI-increasing substitutions (Q311R/D413K) BMS103205H795-MY201, MS103205H795-MY205, MS103205H795, respectively. All MS1032LO06 variants were expressed with M103202L889-SK1 as light chain according to the method described in Example 27 and their affinity for human and cynoFc-gamma R was assessed using the method described in Example 27.

Based on the SPR analysis, the results of which are summarized in Table 24, it was confirmed that pI-increasing substitutions do not affect binding to Fc-gamma R of Fc variants with enhanced Fc-gamma RIIb activity. MY101 and PK2, which were obtained by introducing S400R/D413K into MY101, were found to have 9-fold and 8-fold increased binding to human Fc-gamma RIIb, respectively. The affinity of MY101 and PK2 for human Fc-gamma RIIa remained comparable to SG1. For other human and cynoFc-gamma R, an almost similar binding profile was found. Similarly, MY351 has a similar binding profile to the MY201 parent molecule, MY344 to the MY205 parent molecule, and MY335 to the MY265 parent molecule (Table 21), respectively. These results indicate that any of the pairs of substitutions that increase pI, S400R/D413K or Q311R/D413K, do not affect the affinity for human and cynoFc-gamma R.

PK study conducted in transgenic all human Fc-gamma R mice

In vivo testing using transgenic all human Fc-gamma R mice

The elimination of myostatin and anti-latent myostatin antibody was assessed in vivo after co-administration of anti-latent myostatin antibody and human latent myostatin to all human Fc-gamma R transgenic mice (FIGS. 24 and 25). Antibody to latent myostatin (0.3 mg/ml) and latent myostatin (0.05 mg/ml) were administered in a single dose of 10 ml/kg into the caudal vein. Anti-CD4 antibody (1 mg/ml) was administered three times (every 10 days) at a dose of 10 ml/kg into the caudal vein to suppress anti-drug antibodies. Blood samples were collected at 5 min, 15 min, 1 h, 4 h, 7 h, 1 day, 2 days, 7 days, 14 days, 21 days and 28 days after administration. The collected blood was immediately centrifuged at 15,000 rpm at 4° C. for 5 minutes to separate the plasma. The separated plasma was stored at -20° C. or below until analysis. MS1032LO06-SG1, MS1032LO06-MY101, MS1032LO06-PK2, MS1032LO06-MY201, MS1032LO06-MY351, MS1032LO06-MY205, MS1032LO06-MY344, and MS1032LO06-MY335 were used as antibodies to latent myostatin.

Plasma measurement of total myostatin by electrochemiluminescence (ECL)

The concentration of total myostatin in the plasma of mice was measured using ECL. Anti-mature myostatin antibody immobilized plates were prepared by applying anti-mature myostatin antibody RK35 (described in WO 2009/058346) to a 96-well MULTI-ARRAY plate (Meso Scale Discovery) and incubating in blocking buffer for 2 hours at room temperature. Mature myostatin samples were prepared to obtain a calibration curve and plasma samples diluted 40 times or more. Samples were mixed in an acidic solution (0.2 M glycine-HCl, pH 2.5) to separate by dissociation of mature myostatin from its binding protein (such as a propeptide). The samples were then loaded onto a plate immobilized with anti-mature myostatin antibody and allowed to bind for 1 hour at room temperature, followed by washing. Biotin labeled anti-mature myostatin antibody RK22 (described in WO 2009/058346) was then added and the plate was incubated for 1 hour at room temperature followed by washing. Then, SULFO labeled streptavidin (Meso Scale Discovery) was added and the plate was incubated for 1 hour at room temperature, followed by washing. Read Buffer T (×4) (Meso Scale Discovery) was immediately added to the plate, and signal detection was performed using a SECTOR Imager 2400 (Meso Scale Discovery). Mature myostatin concentration was calculated from the response to the standard curve using SOFTmax PRO analytical software (Molecular Devices). The time dependence of total myostatin plasma concentration after intravenous administration of antibodies to latent myostatin and latent myostatin, determined using this method, is shown in Fig. 24.

Measurement of Plasma Anti-Latent Myostatin Antibody by High Performance Liquid Chromatography-Tandem Electrospray Ionization Spectrometry (LC/ESI-MS/MS)

Anti-latent myostatin antibody concentration in mouse plasma was measured by LC/ESI-MS/MS. Mouse plasma concentrations of calibration standards were 1.5625, 3.125, 6.25, 12.5, 25, 50, 100, and 200 µg/mL. 3 µl of calibration standards and plasma samples were added to 50 µl of Ab-Capture Mag (ProteNova) and allowed to incubate for 2 hours at room temperature. The magnetic beads were then isolated from the samples and washed twice with 0.2 ml (10 mmol/L) PBS with 0.05% Tween 20. The magnetic beads were then washed with PBS (10 mmol/L) to ensure removal of Tween 20. After washing, the magnetic beads were suspended in 25 µl of a mixture containing urea (7.5 mol/l), dithiothreitol (8 mmol/l) and lysozyme (chicken egg protein) (1 µg/ml), 50 mmol/l ammonium bicarbonate, and the suspended samples were incubated for 45 min at 56°C. Then 2 μl of iodoacetamide (500 mmol/l) was added and the samples were incubated for 30 min at 37° C. in the dark. Lysyl endopeptidase digestion was then carried out by adding 150 μl of a solution containing 0.67 μg/ml of lysyl endopeptidase suitable for biochemical analyzes (Wako) in ammonium bicarbonate (50 mmol/l) and the samples were incubated for 3 hours at 37°C. Trypsin digestion was then carried out by adding 10 μl (10 μg/ml) of modified trypsin having a purity suitable for sequencing (Promega) in ammonium bicarbonate (50 mmol/l). Digestion was allowed to take place in the samples by stirring overnight at 37° C. and 5 μl of 10% trifluoroacetic acid was added to terminate the reaction. 50 μl of digested samples were analyzed by LC /ESI-MS/MS. LC/ESI-MS/MC was performed using a Xevo TQ-S triple quadrupole device (Waters) equipped with 2D-UPLC class I (Waters). Anti-latent myostatin specific peptide YAFGQGTK and lysozyme specific peptide GTDVQAWIR used as an internal standard were monitored using Selected Reaction Monitoring (SRM) mode. The SRM transition was [M+2H]2+ (m/z 436.2) to y8 ion (m/z 637.3) for latent myostatin antibody and [M+2H]2+ (m/z 523, 3) to the y8 ion (m/z 545.3) for lysozyme. An internal standard curve was created using a weighted (1/× or 1/×2) linear regression using the dependence of peak area on concentrations. The plasma concentration in mice was calculated using a calibration curve using Masslynx analytical software, version 4.1 (Waters). The time dependence of plasma antibody concentration after intravenous administration of anti-latent myostatin antibody and latent myostatin measured by this method is shown in FIG. 25.

Effect of pI and Fc-gamma R binding on myostatin concentration in vivo

After the introduction of MS1032LO06-SG1, the concentration of total myostatin in plasma after 7 hours decreased by 5 times compared with the concentration of total myostatin in plasma after 5 minutes. In contrast, after administration of MS1032LO06-MY101, MS1032LO06-MY201, and MS1032LO06-MY205, plasma total myostatin concentration after 7 hours decreased by 28-200 times compared with plasma total myostatin concentration after 5 minutes. In addition, after the administration of MS1032LO06-PK2, MS1032LO06-MY351, MS1032LO06-MY344, and MS1032LO06-MY335, the concentration of total myostatin in plasma after 7 hours decreased by 361-419 times compared with the concentration of total myostatin in plasma after 5 minutes. On the other hand, the difference in the concentration of each antibody at each time point tested compared to MS1032LO06-SG1 was found to be approximately 2-fold, and the pI variants did not increase antibody elimination from plasma. Substitutions resulting in both increased binding to human Fc-gamma RIIb and increased pI increased myostatin but not antibody elimination.

Variants with high pI had a greater positive charge in plasma. Therefore, this positive charge interacted with the negatively charged cell surface, the high pI variants antigen-antibody immune complex contacted the cell surface more strongly, resulting in increased uptake of the high pI variants antigen-antibody immune complex by the cell.

Example 29

Evaluation of myostatin clearance with Fc variants with increased activity against Fc-gamma RIIb in monkeys

The effect of increased binding capacity of Fc variants generated according to the method described in Example 27 to cynoFc-gamma RIIb on myostatin elimination was evaluated in cynomolgus monkeys. In addition, the combined effect with substitutions leading to an increase in pI was also evaluated.

Obtaining and profiling scoring options

The 14 antibodies tested and their binding profiles are summarized in Table 25. The presence of an Fc variant with increased ability to bind to FcRn at acidic pH has been reported to increase the in vivo half-life of the antibody (J. Biol. Chem. 281, 2006, pp. 23514- 23524; Nat Biotechnol 28, 2010, pp. 157-159, Clin Pharm. & Thera 89(2), 2011, pp. 283-290). To increase the half-life of the antibody without binding to rheumatoid factor, the invention combined these substitutions with Fc variants with increased activity against Fc-gamma RIIb and increased pI.

Тяжелые цепи MS103205H795-SG1012, MS103205H795-SG1029, MS103205 H795-SG1031, MS103205H795-SG1033, MS103205H795-SG1034 получали путем интродукции замены N434A в MS103205H795-MY101, MS103205 H795-MY344, MS103205H795-MY351, MS103205H795-MY201, MS103205H795-MY335, соответственно . MS103205H795-SG1016 was generated by introducing pI-increasing substitutions (Q311R/D399K) into MS103205H795-SG1012. MS103240H795-SG1071 and MS103240H795-SG1079 were obtained by introducing the M428L/N434A/Y436T/Q438R/S440E substitutions and the N434A/Q438R/S440E substitutions into MS103240H795-MY344, respectively (MS103240H795: 92 SEQ ID).

MS103240H795-SG1074 and M8103240H795-SG1077 were generated by introducing the pI-increasing Q311R/P343R and M428L/N434A/Y436T/Q438R/S440E substitutions in MS103240H795-MY209 and MS103240H795-MY5895, respectively. MS103240H795-SG1080 and MS103240H795-SG1081 were generated by introducing pI-increasing substitutions Q311R/P343R and substitutions N434A/Q438R/S440E in MS103240H795-MY209 and MS103240H795-MY518, respectively. MS103240H795-SG1071 was generated by introducing pI-increasing substitutions Q311R/D413K and substitutions M428L/N434A/Y436T/Q438R/S440E in MS103240H795-MY205. MS103240H795-SG1079 was generated by introducing pI-increasing substitutions Q311R/D413K and substitutions N434A/Q438R/S440E in MS103240H795-MY205. These MS1032LO06 variants and MS1032LO19 variants were expressed with M103202L889-SK1 and M103202L1045-SK1 respectively (M103202L1045: VL, SEQ ID NO: 97) as light chain according to the method described in Example 34, and their affinity for human and cynoFc-gamma R was evaluated using the method described in Example 27. In this example, the fold KD value for each Fc variant was calculated by dividing the KD value for the parent SG1 molecule by the KD value for the variant for each Fc-gamma R.

For example, the multiples of KD values for MS1032LO06-SG1012 and MS1032LO19-SG1071 were calculated by dividing the KD values for MS1032LO06-SG1 and MS1032LO19-SG1 by the KD values for MS1032LO06-SG1012 and MS1032LO19-SG1071, respectively.

Table 25 summarizes the results of the SPR analysis. Of these variants, SG1012, SG1016, SG1074, and SG1080 showed the strongest affinity for human Fc-gamma RIIb, with 10-fold higher affinity for human Fc-gamma RIIb compared to SG1.

29.2. PK study conducted on monkeys

In vivo testing using cynomolgus monkeys

The accumulation of endogenous myostatin was assessed in vivo after administration of antibodies to latent myostatin to 2-4-year-old Masaca fascicularis (cynomolgus cynomolgus monkey) from Cambodia (Shin Nippon Biomedical Laboratories Ltd., Japan). A dose of 30 mg/kg was injected into the cephalic vein of the forearm using a disposable syringe, an extension tube, an internal needle and an infusion pump. The dosing rate was 30 minutes per animal. Blood samples were collected before the start of dosing and either 5 min, 7 h and 1, 2, 3, 7, 14, 21, 28, 35, 42, 49 and 56 days after the end of dosing, or 5 min and 2, 4 and 7 hours, and 1, 2, 3, 7, 14, 21, 28, 35, 42, 49 and 56 days after the end of dosing. Blood was taken from the femoral vein using a syringe containing sodium heparin. The blood was immediately cooled on ice and plasma was obtained by centrifugation at 4°C, 1700×g for 10 min. Plasma samples were stored in a deep freezer (acceptable range: -70°C or below) until analysis. В качестве антител к латентному миостатину применяли MS1032LO00-SG1, MS1032LO06-SG1012, MS1032LO06-SG1016, MS1032LO06-SG1029, MS1032LO06-SG1031, MS1032LO06-SG1033 и MS1032LO06-SG1034 (в контексте настоящего описания SG1012, SG1016, SG1029, SG1031, SG1033 и SG1034 denote heavy chain constant regions constructed from SG1 according to the method described below).

Anti-latent myostatin antibodies MS1032LO19-SG1079, MS1032LO19-SG1071, MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081 and MS1032LO19-SG1081 and (as used herein, SG1079, SG1071, SG1080, SG1074, SG1081, and SG1077 denote heavy chain constant regions constructed from SG1 according to the method described below). Blood samples were collected before the start of dosing and 5 minutes and 2, 4 and 7 hours, and 1, 2, 3, 7, 14 days after the end of dosing. The blood was processed according to the procedure described above. Plasma samples were stored in a deep freezer (acceptable range: -70° C. or below) until analysis.

Plasma measurement of total myostatin by electrochemiluminescence (ECL)

Plasma total myostatin concentration in monkeys was measured by ECL according to the method described in Example 23. The time course of plasma total myostatin concentration after intravenous administration of anti-latent myostatin antibody, determined using this method, is shown in FIG. 26.

Measurement of monkey plasma ADA by electrochemiluminescence (ECL)

The biotinylated drug was used to sensitize a 96-well MULTI-ARRAY streptavidin plate (Meso Scale Discovery) and incubated in LowCross® buffer (Candor) for 2 hours at room temperature. Monkey plasma samples were diluted 20-fold in LowCross® buffer prior to loading into the plate. Samples were incubated overnight at 4°C. The next day, the plate was washed three times with wash buffer before adding a SULFO-labeled anti-monkey IgG secondary antibody (Thermo Fisher Scientific). After incubation for 1 hour at room temperature, the plate was washed three times with wash buffer. Read Buffer T (x4) (Meso Scale Discovery) was immediately added to the plate, and signal detection was performed with a SECTOR Imager 2400 (Meso Scale Discovery).

Effect of pH Dependence and Fc Engineering on Myostatin Accumulation in Monkeys in Vivo

Administration of pH-independent antibody (MS1032LO00-SG1) to cynomolgus monkeys resulted in at least a 60-fold increase in myostatin concentration on day 28 from baseline. On day 28, pH-dependent latent myostatin antibodies MS1032LO06-SG1012 and MS1032LO06-SG1033 provided 3- and 8-fold increases from baseline, respectively. The strong sweeping effect is mainly associated with an increase in affinity for Fc-gamma RIIb of cynomolgus monkeys. pH-dependent antibodies to latent myostatin MS1032LO06-SG1029, MS1032LO06-SG1031 and MS1032LO06-SG1034 were able to eliminate the antigen by day 28 to a level below baseline. The reason for the strong sweeping effect induced by MS1032LO06-SG1029, MS1032LO06-SG1031 and MS1032LO06-SG1034IO) is an increase in non-specific cellular uptake due to an increase in the positively charged antibody cluster and an increase in Fc-gamma R mediated cellular uptake due to increased Fc-binding. gamma R.

The introduction of pH-dependent antibodies to latent myostatin MS1032LO19-SG1079, MS1032LO19-SG1071, MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081 and MS1032LO19-SG1077 reduced the concentration of myostatin to a level below the detection limit (<0.25 ng/mL), starting from from day 1 in cynomolgus monkeys. On day 14, myostatin concentrations exceeded the detection limit with MS1032LO19-SG1079 and MS1032LO19-SG1071, although myostatin levels remained below the detection limit with MS1032LO19-SG1080, MS1032LO19-SG1074, MS1032LO19-SG1081, and MS1032LO19-SG1077. The weaker suppression found for MS1032LO19-SG1079 and MS1032LO19-SG1071 may be the result of various mutations leading to a change in pI.

The results suggest that it is possible to achieve a strong elimination of myostatin from plasma using mutations that increase binding to Fc-gamma RIIb or combinations of mutations that increase the positive charge of the antibody and increase binding to Fc-gamma RIIb. It can be expected that a strong elimination of myostatin can be achieved in humans by combining mutations that increase the positive charge of the antibody and increase binding to Fc-gamma R.

Example 30

Screening for pI-increasing substitutions to increase myostatin clearance

In this example, the role of pI-increasing substitutions in the Fc region of an antibody in increasing myostatin clearance was evaluated. The method of introducing amino acid substitutions in the constant region of an antibody to increase pI is not limited to the above method, but for example, it can be carried out according to the method described in WO 2014/145159. As with the variable region, the amino acid substitutions that were introduced into the constant region were preferably substitutions that reduced negatively charged amino acids (such as aspartic acid and glutamic acid) while increasing positively charged amino acids (such as arginine and lysine). In addition, amino acid substitutions can be introduced at any position in the antibody constant region and can be a single amino acid substitution or combinations of several amino acid substitutions. The sites of introduction of amino acid substitutions are preferably, but not limited to, positions at which amino acid side chains can be exposed on the surface of the antibody molecule. Most preferred examples include the method of introducing a combination of several amino acid substitutions at positions that can be exposed on the surface of an antibody molecule. Alternatively, the multiple amino acid substitutions introduced in the course of the present invention are preferably positioned so that they are in structural proximity to each other. In addition (but not limited to), several of the amino acid substitutions introduced during the creation of the present invention, preferably were substitutions for positively charged amino acids, so that it preferably leads to a state in which many positive charges are present in structurally similar positions.

Obtaining and profiling studied variants

The antibodies studied are summarized in Table 26. MS103205H795-SG141 heavy chain was generated by introducing pI-increasing Q311R/D399R substitutions in MS103205H795-SG1. Other heavy chain variants were also made by introducing the appropriate substitutions shown in Table 26 in MS103205H795-SG1. All variants of MS1032LO06 were expressed with M103202L889-SK1 as light chain according to the method described in Example 34.

Evaluation of binding activity to mouse Fc-gamma RII Fc variants with increased pI using BIACORE®

Whether or not variant-containing Fc-region antibodies were obtained, binding assays between soluble mouse Fc-gamma RII and antigen-antibody complexes were performed using BIACORE® T200 (GE Healthcare). Soluble mouse Fc-gamma RII was obtained in the form of a His labeled molecule using a method known to those skilled in the art. An appropriate amount of anti-His antibody was fixed on a CM5 sensor chip (GE Healthcare) by the amine coupling method using a His capture kit (GE Healthcare) to immobilize mouse Fc-gamma RII. The antibody-antigen complex and running buffer (as reference solution) were then injected and allowed to react with mouse Fc-gamma RII immobilized on the sensor chip. 20 mM N-[-(2-acetamido)-2-aminoethanesulfonic acid, 150 mM NaCl, 1.2 mM CaCl ₂ and 0.05% (w/v) Tween 20 pH 7.4 were added as mobile buffer, and an appropriate buffer was also used to dilute soluble mouse Fc-gamma RII. To regenerate the sensor chip, 10 mM glycine-HCl, pH 1.5 was used. All measurements were carried out at 25°C. The analyzes were performed based on the binding (RU) calculated from the sensograms obtained by measurements, and relative values are presented, for which the binding level of SG1 was taken as 1.00. Parameters were calculated using Evaluation BIACORE® T100 software (GE Healthcare).

The results of the SPR analysis are summarized in Table 26. Several Fc variants were found to have increased affinity for mouse Fc-gamma RII fixed on the BIACORE® sensor chip.

Without being limited by any particular theory, this result can be explained as follows. As is known, the BIACORE® sensor chip is negatively charged, and it can be assumed that such a charged state resembles the state of the cell membrane surface. More specifically, the affinity of the antigen-antibody complex for a mouse Fc-gamma RII fixed on a negatively charged BIACORE® sensor chip would likely resemble the binding of an antigen-antibody complex to a mouse Fc-gamma RII present on a similarly negatively charged cell membrane surface.

As used herein, antibodies obtained by introducing pI-enhancing modifications into the Fc region are antibodies in which the charge of the Fc region is more positive than before the introduction of the modifications. Thus, it can be assumed that the Coulomb interaction between the Fc region (positive charge) and the surface of the sensor chip (negative charge) will be enhanced when using amino acid modifications that increase pI. In addition, similar phenomena are expected to occur on the same negatively charged surface of the cell membrane; therefore, it can also be expected that they reflect the phenomena of increasing the rate of uptake by the cell in vivo.

Of the Fc variants with elevated pI with two amino acid substitutions relative to SG1, the antigen-antibody complex formed with SG141, P1499m, P1501m and P1540m exhibited the highest level of binding to human Fc-gamma RIIb. The amino acid substitutions Q311R/D399R, Q311R/P343R, Q311R/D413R and D401R/D413K were confirmed to have a strong charge-dependent effect on binding to human Fc-gamma RIIb on the sensor chip.

Cellular uptake of Fc variants with increased pI

To evaluate the rate of intracellular uptake of a human Fc-gamma RIIb cell line expressing, the following assay was performed. MDCK (Maidin-Derby dog kidney cell line) cells that constitutively express human Fc-gamma RIIb were obtained by known methods. These cells were used to evaluate intracellular uptake of antigen-antibody complexes.

In particular, pHrodoRed (Life Technlogies) was used to label human latent myostatin (antigen) according to a known protocol, and antigen-antibody complexes were obtained in culture solution with an antibody concentration of 10 mg/ml and an antigen concentration of 2.5 mg/ml. The culture solution containing the antigen-antibody complexes was added to culture plates with the above MDCK cells that constitutively express human Fc-gamma RIIb and incubated for 1 hour, and then the fluorescence intensity of the antigen taken up by the cells was quantified using the InCell analyzer 6000 (GE healthcare). The amount of absorbed antigen was expressed as relative values, while the value for SG1 was taken as 1.00.

The quantitative results of cellular uptake are summarized in Table 26.

When using some variants of Fc found a strong fluorescence in the cells inherent in the antigen.

Without being limited by any particular theory, this result can be explained as follows.

The antigen and antibodies added to the cell culture solution form antigen-antibody complexes in the culture solution. The antigen-antibody complexes bind to the human Fc-gamma RIIb expressed on the cell membrane via the Fc region of the antibody and are taken up by cells in a receptor-dependent manner. The antibodies used in this experiment bind to the antigen in a pH dependent manner; thus, the antibody can dissociate from the antigen in endosomes (acidic pH conditions) within cells. Since the dissociated antigen was labeled with pHrodoRed as described earlier, its fluorescence occurs in endosomes. Thus, the strong intensity of fluorescence inside the cell probably indicates that the absorption of antigen-antibody complexes into cells occurred at a faster rate or in greater quantities.

Among the Fc variants with increased pI with two amino acid substitutions relative to SG1, antigen-antibody complexes formed from SG141, P1375m, P1378m, P1499m, P1524m, P1525m, P1532m, P1533m, P1540m, P1541m, and P1543m showed higher uptake of antigen by cells. It was confirmed that amino acid substitutions Q311R/D399R, Q311R/D413K, S400R/D413K, Q311R/P343R, Q311R/G341R, Q311R/G341K, Q311R/D401R, Q311R/D401K, D401R/D413K, D401K/D413 charge-mediated effect on cellular uptake of the antigen-antibody complex.

PK study conducted in transgenic human FcRn mice

In vivo testing using transgenic human FcRn mice

Elimination of myostatin and anti-latent myostatin antibody was assessed in vivo after co-administration of anti-latent myostatin antibody and latent myostatin to human FcRn transgenic mice. Anti-latent myostatin antibody (0.1 mg/ml) and mouse latent myostatin (0.05 mg/ml) were administered at a single dose of 10 ml/kg into the caudal vein in experiment PK-2 (described in FIG. 27). Antibody to latent myostatin (0.3 mg/ml), human latent myostatin (0.05 mg/ml) and human normal immunoglobulin (CSL Behring AG) (100 mg/ml) were administered in a single dose of 10 ml/kg into the caudal vein in the PK-4 experiment (described in Fig. 28). Blood samples were collected at 5 min, 15 min, 1 h, 4 h, 7 h, 1 day, 2 days, 7 days, 14 days, 21 days and 28 days after administration in the PK-2 experiment. Blood samples were collected at 5 min, 1 h, 4 h, 7 h, 1 day, 7 days, 14 days, 21 days and 30 days after administration in experiment PK-4. The collected blood was immediately centrifuged at 15,000 rpm at 4° C. for 5 minutes to separate the plasma. The separated plasma was stored at -20° C. or below until analysis. Antibodies to latent myostatin MS1032LO06-SG1, MS1032LO06-P1375m, MS1032LO06-P1378m, MS 1032LO06-P1383m were used in the PK-2 experiment, and MS1032LO06-P1375m, MS1032LO06-P1499m were used in the PK-4 experiment.

Plasma measurement of total myostatin by electrochemiluminescence (ECL)

The concentration of total myostatin in the plasma of mice was measured using ECL according to the method described in example 28 (Ravetch PK). Data on the time dependence of the concentration of total myostatin in plasma after intravenous administration of antibodies to latent myostatin and latent myostatin, obtained using this method, are presented in Fig. 27A and 28A.

Measurement of the concentration of antibodies to latent myostatin in plasma using enzyme-linked immunosorbent assay (ELISA)

Anti-latent myostatin antibody concentration in mouse plasma was measured by ELISA in the PK-2 experiment. Anti-human IgG (gamma chain specific) F(ab')2 fragment (Sigma) was applied to a Nunc-Immuno MaxiSorp plate (Nalge Nunc International) and allowed to stand overnight at 4°C to prepare plates with immobilized anti-human IgG. Plasma concentrations of 2.5, 1.25, 0.625, 0.313, 0.156, 0.078, and 0.039 μg/mL and mouse plasma samples diluted 100-fold or more were prepared. Samples were then plated onto immobilized anti-human IgG plates and allowed to stand for 1 hour at room temperature. Goat anti-human IgG (gamma chain specific)-biotin conjugate (Southern Biotech) was then added and allowed to react for 1 hour at room temperature. Streptavidin-PolyHRP80 (Stereospecific Detection Technologies) was then added and allowed to react for 1 hour at room temperature and a chromogenic reaction was performed using TMB One Component HRP Microwell (BioFX Laboratories) as the substrate. After termination of the reaction with 1 N. sulfuric acid (Showa Chemical) absorbance at 450 nm was measured using a microplate reader. The plasma concentration in mice was calculated from the absorbance of the calibration curve using SOFTmax PRO analytical software (Molecular Devices). Data on the time dependence of the concentration of antibodies in plasma after intravenous administration of antibodies to latent myostatin and latent myostatin, obtained by this method, are presented in Fig. 27B.

In the PK-4 experiment, the plasma concentration of anti-latent myostatin antibody in mice was measured using the Gyrolab Bioaffy™ CD method (Gyros). The biotinylated anti-human IgG-Fc antibody was passed through a column of streptavidin beads in a CD microstructure. Samples were prepared for the calibration curve at plasma concentrations of 0.5, 1, 2, 4, 8, 16 and 32 μg/ml by injecting human normal immunoglobulin (CSL Behring AG) at a concentration of 5 mg/ml and at a concentration of 200 μg/ml mouse latent myostatin, and mouse plasma samples were diluted 25-fold or more by injecting into plasma at a concentration of 200 μg/ml mouse latent myostatin. Samples were then added to CD and passed through a bead column. Then, an Alexa-labeled goat anti-human IgG polyclonal antibody (BETHYL) was added to the CD and passed through a bead column. Mouse plasma concentration was calculated from the calibration curve using the Gyrolab Evaluator software. Data on the time dependence of the concentration of antibodies in plasma after intravenous administration of antibodies to latent myostatin and latent myostatin, obtained by this method, are presented in Fig. 28B.

Effect of pI on myostatin concentration in vivo

After the introduction of MS1032LO06-SG1, the plasma concentration of total myostatin after 7 hours decreased by 12 times compared with the plasma concentration of total myostatin after 5 minutes in the PK-2 experiment. In contrast, after administration of MS 1032LO06-P 1375m, MS 1032LO06-P1378m, and MS 1032LO06-P1383m, the plasma concentration of total myostatin after 7 h decreased by 26-67 times compared with the plasma concentration of total myostatin after 5 min in the PK-2 experiment. . The concentration of antibodies MS 1032LO06-P 1378m and MS 1032LO06-P1383m decreased by more than 3 times compared to MS1032LO06-SG1, although the differences in the concentration of MS1032LO06-P1375m at each tested time point compared to MS1032LO06-SG1 turned out to be approximately 2-fold in the experiment PK-2. In the presence of pI variants, including amino acid substitutions Q311R/D413K, an increased elimination of myostatin from plasma was found without an increase in the elimination of antibody from plasma.

In addition, total myostatin concentration and antibody concentration were determined after administration of MS 1032LO06-P1499m by co-administration of human normal immunoglobulin to mimic human plasma. After the introduction of MS1032LO06-P1375m and MS1032LO06-P1499m, the concentration of total myostatin in plasma after 7 hours decreased by 3.4 and 5.3 times compared with the concentration of total myostatin in plasma after 5 minutes, and the concentration of total myostatin in plasma and the concentration of antibodies in each the tested time point when using MS 1032LO06-P1499m was 1.5 times higher than those established for MS 1032LO06-P1375m in the PK-4 experiment. In the presence of pi variants, including amino acid substitutions Q311R/P343R, an increased elimination of myostatin from plasma was found without an increase in the elimination of antibody from plasma.

Variants with high pI had a greater positive charge in plasma. Therefore, this positive charge interacted with the negatively charged cell surface, the antigen-antibody immune complex of the high pI variants contacted the cell surface more strongly, resulting in increased uptake of the antigen-antibody complex of the high pI variants by the cell.

Example 31

Substitutions leading to an increase in pI in combination with an Fc variant with increased activity against Fc-gamma RIIb

The effect on myostatin clearance of an Fc variant with increased activity against Fc-gamma RIIb in combination with other pI-increasing substitutions was evaluated in transgenic human Fc-gamma RIIb mice. Human Fc-gamma RIIb transgenic mice were generated by microinjection of a BAC (bacterial artificial chromosome) vector containing exons of the human FCGR2B gene into the pronucleus of fertilized eggs of C57BL/6N mice using standard techniques (see, for example, J. Immunol., 195 (7), October 1, 2015, pp. 3198-3205). Mouse Fc-gamma RIIb deficient mice were generated using zinc finger nucleases (ZFNs) that were designed to target exon 1. Humanized Fc-gamma RIIb mice were generated by crossing human Fc-gamma RIIb transgenic mice with mice with "off" (KO) mouse Fc-gamma RIIb. Using these mice, the combined effects of increased affinity for human Fc-gamma RIIb and increased pI on soluble antigen clearance can be studied.

Obtaining and profiling test cases

The 7 antibodies tested and their binding profiles are summarized in Table 27. The heavy chain, MS103205H795-MY352, MS103205H795-PK55, MS103205H795-PK56, MS103205H795-PK57, was generated by introducing pI-increasing substitutions Q311R/D411R3R, Q311R, Q311R /P343R/D413R, Q311R/N384R/D413R, respectively in MS103205H795-MY201. All MS1032LO06 variants were expressed using M103202L889-SK1 as light chain according to the method described in Example 30 and their affinity for human and cynoFc-gamma R was determined using the method described in Example 27.

Based on the results of the SPR analysis, which are summarized in Table 27, it was confirmed that substitutions resulting in an increase in pI did not affect binding to Fc-gamma R of Fc variants with increased activity against Fc-gamma RIIb. MY201 and MY351, which were obtained by introducing Q311R/D413K into MY201, were found to have 6-fold and 5-fold increased binding to human Fc-gamma RIIb, respectively. They had almost the same binding profile with other human and cynoFc-gamma R. Similarly, MY352, PK55, PK56 and PK57 showed a similar binding profile to that of the parent MY201 molecule (Table 27). These results indicate that any pair of substitutions resulting in an increase in pI did not affect the affinity for human and cynoFc-gamma R.

PK study conducted in transgenic human FcRn mice

In vivo testing using transgenic human FcRn mice

The elimination of myostatin and anti-latent myostatin antibody was assessed in vivo after co-administration of anti-latent myostatin antibody and human latent myostatin to human Fc-gamma RIIb transgenic mice in which mouse Fc-gamma RII was replaced with human Fc-gamma RIIb. Antibody to latent myostatin (0.3 mg/ml) and mouse latent myostatin (0.05 mg/ml) were administered in a single dose of 10 ml/kg into the caudal vein. Blood samples were collected at 5 min, 1 h, 4 h, 7 h, 1 day, 7 days, 14 days, 21 days and 28 days after administration. The collected blood was immediately centrifuged at 15,000 rpm at 4° C. for 5 minutes to separate the plasma. The separated plasma was stored at -20° C. or below until analysis. The following latent myostatin antibodies MS1032LO06-SG1, MS1032LO06-MY201, MS1032LO06-MY351, MS1032LO06-MY352, MS1032LO06-PK55, MS1032LO06-PK56 and MS1032LO06-PK57 were used.

Plasma measurement of total myostatin by electrochemiluminescence (ECL)

The plasma concentration of total myostatin in mice was measured by ECL according to the method described in Example 28. The time-dependence data of the plasma concentration of total myostatin after intravenous administration of anti-latent myostatin antibody and latent myostatin obtained using this method are shown in FIG. 29.

Measurement of the concentration of antibodies to latent myostatin in plasma using enzyme-linked immunosorbent assay (ELISA)

The concentration of antibodies to latent myostatin in the plasma of mice was measured using ELISA. Anti-human IgG (gamma chain specific) F(ab')2 fragment (Sigma) was applied to a Nunc-Immuno MaxiSorp plate (Nalge Nunc International) and allowed to stand overnight at 4°C to obtain plates with immobilized anti-human IgG. Plasma concentrations of 2.5, 1.25, 0.625, 0.313, 0.156, 0.078, and 0.039 μg/mL and mouse plasma samples diluted 100-fold or more were prepared. Samples were then plated onto immobilized anti-human IgG plates and allowed to stand for 1 hour at room temperature. Goat anti-human IgG (gamma chain specific)-biotin conjugate (Southern Biotech) was then added and allowed to react for 1 hour at room temperature. Streptavidin-PolyHRP80 (Stereospecific Detection Technologies) was then added and allowed to react for 1 hour at room temperature and a chromogenic reaction was performed using TMB One Component HRP Microwell (BioFX Laboratories) as the substrate. After termination of the reaction with 1 N. sulfuric acid (Showa Chemical) absorbance was measured at 450 nm using a microplate reader. The plasma concentration in mice was calculated from the absorbance of the calibration curve using SOFTmax PRO analytical software (Molecular Devices). Data on the time dependence of the concentration of antibodies in plasma after intravenous administration of antibodies to latent myostatin and latent myostatin, obtained by this method, are presented in Fig. thirty.

Effect of pI and Fc-gamma R-binding on myostatin concentration in vivo

After the introduction of MS1032LO06-MY201, the plasma concentration of total myostatin after 7 hours decreased by 8 times compared with the plasma concentration of total myostatin after 5 minutes. In contrast, after administration of MS1032LO06-PK57, the plasma concentration of total myostatin after 7 hours decreased by 376 times compared with the plasma concentration of total myostatin after 5 minutes. For other variants with high pI, a similar increase in the elimination of myostatin from plasma was found. The concentration of high pI antibody variants after 28 days decreased by 2-3 times compared with the concentration of the MS1032LO06-SG1 antibody, and analysis for pI variants revealed a slight increase in the elimination of antibodies from plasma.

Variants with high pI had a greater positive charge in plasma. Therefore, this positive charge interacted with the negatively charged cell surface, the antigen-antibody immune complex of the high pI variants contacted the cell surface more strongly, resulting in increased uptake of the antigen-antibody complex of the high pI variants by the cell.

Example 32

Creation of Fc variants with enhanced activity against human Fc-gamma RIIb

Fc variants with enhanced activity against human Fc-gamma RIIb having an anti-myostatin antibody variable region were constructed and their affinity for human Fc-gamma R was evaluated. In particular, Fc variants with increased activity against human Fc-gamma RIIb were constructed by introducing substitutions in MS103240H795-SG1 (VH, SEQ ID NO: 92; CH, SEQ ID NO: 9) in combination with substitutions T250V/T307P. In addition, substitutions (Q311R/D413K or Q311R/P343R) were also introduced. Variants were expressed using M103202L1045-SK1 as light chain and their affinity for human Fc-gamma R was analyzed according to the method described in Example 34. Table 28 shows the results of kinetic analyzes for human and cynoFc-gamma R. KD values that indicated in the cells of the table in bold were calculated using equation 2, because their affinity was too low to accurately determine it using kinetic analysis.

All studied variants showed increased affinity for human Fc-gamma RIIb and reduced ability to bind to human Fc-gamma RIIaR, Fc-gamma RIIaH, Fc-gamma RIIIaV compared to SG1. The affinity for human Fc-gamma RIIb ranged from 4.1-fold to 30.5-fold compared to SG1. The affinity for human Fc-gamma RIIaR was between 0.02-fold and 0.22-fold. Affinities for human Fc-gamma RIIaH and human Fc-gamma RIIIaV were less than 0.04-fold and 0.012-fold, respectively. It was found that the substitutions (T250V/T307P) did not affect the binding profile to human Fc-gamma R compared to the affinity of MY009 (P238D) and MY214 (P238D/T250V/T307P). On the other hand, both pairs of pI-increasing substitutions (Q311R/D413K and Q311R/P343R) reduced binding affinity by about 0.5-fold compared to variants that did not contain pI-increasing substitutions. For example, although TT14 was found to have a 30.5-fold higher affinity for human Fc-gamma RIIb, only 16.1-fold and 145-fold increases were found when used in combination with Q311R/D413K (TT33) and Q311R/P343R (TT32 ) respectively.

In addition, the substitutions described in Example 29, which extended the half-life of the antibody and reduced binding to rheumatoid factor, were introduced into Fc variants with increased activity against human Fc-gamma RIIb and analyzed for their affinity for human Fc-gamma R (table 29). In these SPR assays, all data were obtained using mouse anti-human kappa light chain IgG to "capture" the antibodies of interest. Table 30 shows the results, while the KD values in bold cells were calculated using Equation 2 because their affinity was too low to be accurately determined by kinetic analysis.

In this experiment, SG1 and TT33 were re-evaluated, with a “capture” method different from the one presented in Table 28. As a result, it was found that the KD values for TT33 corresponded to those obtained using the method, the results of which are shown in Table 28 (16.1-fold level in table 28 and 15.7-fold level in table 30 characterizing binding to human Fc-gamma RIIb). For TT92 and TT93, which were constructed by introducing N434A/Q438R/S440E and M428L/N434A/Y436T/Q438R/S440E into TT33, a binding profile comparable to TT33 was found. Similarly, for TT90 and TT91, which were obtained from TT32, TT72 and TT73, which were obtained from TT31, TT70 and TT71, which were obtained from TT30, TT68 and TT69, which were obtained from TT21, and TT66 and TT67, which were obtained from TT20, found a binding profile comparable to the parental antibodies shown in Table 28. These results indicate that substitutions designed to increase the half-life of the antibody and reduce binding to rheumatoid factor do not affect the binding profile to human Fc-gamma R Fc variants with increased activity against human Fc-gamma RIIb.

Example 33

Cell Imaging Assay of Fc Variants with Enhanced Activity on Human Fc-gamma RIIb

To study the intracellular uptake of an antigen-antibody complex formed with the antibodies of Example 32, the cell imaging assay described in Example 30 was performed. was 1.25 mg/ml and the antigen concentration was 0.34 mg/ml.

The results of the analysis are shown in Fig. 31. With increasing binding affinity to human Fc-gamma RIIb, uptake of antigen-antibody complex by cells increased. In the case of TT14, which has a 30-fold increased binding affinity for human Fc-gamma RIIb compared to SG1, the fluorescence intensity of the antigen inside the cells was 7.5 times higher than the fluorescence intensity of SG1.

Although pI-increasing substitutions (Q311R/D413K and Q311R/P343R) reduced binding affinity for human Fc-gamma RIIb by about 0.5-fold compared to variants that did not contain pI-increasing substitutions, cellular uptake of the antigen-antibody complex with Fc variants containing pI-increasing substitutions increased compared to Fc variants without pI-increasing substitutions. For TT33, a 36-fold increase in cellular uptake of the antigen-antibody complex was found compared to SG1, while for the parent molecule TT14, a 7.5-fold increase in cellular uptake was found. In addition, uptake of the antigen-antibody complex by cells was found for TT33, comparable to the uptake by cells of the SG1071, SG1074, SG1077, SG1079, SG1080, and SG1081 variants, for which a strong sweeping effect was found when studying cynomolgus monkeys. These results suggest that combining pI-increasing substitutions with Fc variants with increased activity against Fc-gamma RIIb is an effective tool for eliminating antigens.

Example 34

Construction of antibody expression vectors and expression and purification of antibodies

Synthesis of full-length genes encoding the nucleotide sequences of the H chain and L chain of antibody variable regions was carried out using methods of preparation known to those skilled in the art, using PCR used for assembly, and the like. The introduction of amino acid substitutions was carried out according to methods known to those skilled in the art using PCR or the like. The resulting plasmid fragments were inserted into an animal cell expression vector, and an H chain expression vector and an L chain expression vector were obtained. The nucleotide sequence of the resulting expression vector was determined using methods known to those skilled in the art. The resulting plasmids were briefly introduced into the HEK293H cell line derived from human embryonic kidney cancer cells (Invitrogen) or FreeStyle293 cells (Invitrogen) for antibody expression. The resulting culture supernatant was collected and passed through a 0.22 µm MILLEX®-GV filter (Millipore) or through a 0.45 µm MILLEX®-GV filter (Millipore) to obtain a culture supernatant. Antibodies were purified from the obtained culture supernatant using methods known to those skilled in the art using r(recombinant) Protein A-Sepharose Fast Flow (GE Healthcare) or Protein G-Sepharose 4 Fast Flow (GE Healthcare). To determine the concentration of purified antibodies, their absorbance was measured at 280 nm using a spectrophotometer. From these values, the antibody concentration was determined using an extinction coefficient calculated using methods such as PACE (Protein Science 4, 1995, pp. 2411-2423).

Although the invention has been described above for a better understanding with some details for the purpose of illustration and examples, the description and examples should not be construed as limiting the scope of the invention. The description of all patent and scientific literature cited in the present description is expressly incorporated herein by reference in its entirety.

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SEQUENCE LIST

<110> CHUGAI SEYYAKU KABUSHIKI KAISHA

<120> Myostatin antibodies, polypeptides containing variants

Fc-regions and how to use them

<130> C1-A1518P

<150> JP 2015-247070

<151> 2015-12-18

<160> 381

<170>PatentIn version 3.5

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Met Gln Lys Leu Gln Leu Cys Val Tyr Ile Tyr Leu Phe Met Leu Ile

1 5 10 15

Val Ala Gly Pro Val Asp Leu Asn Glu Asn Ser Glu Gln Lys Glu Asn

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Val Glu Lys Glu Gly Leu Cys Asn Ala Cys Thr Trp Arg Gln Asn Thr

35 40 45

Lys Ser Ser Arg Ile Glu Ala Ile Lys Ile Gln Ile Leu Ser Lys Leu

50 55 60

Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Asp Val Ile Arg Gln Leu

65 70 75 80

Leu Pro Lys Ala Pro Pro Leu Arg Glu Leu Ile Asp Gln Tyr Asp Val

85 90 95

Gln Arg Asp Asp Ser Ser Asp Gly Ser Leu Glu Asp Asp Asp Tyr His

100 105 110

Ala Thr Thr Glu Thr Ile Ile Thr Met Pro Thr Glu Ser Asp Phe Leu

115 120 125

Met Gln Val Asp Gly Lys Pro Lys Cys Cys Phe Phe Lys Phe Ser Ser

130 135 140

Lys Ile Gln Tyr Asn Lys Val Val Lys Ala Gln Leu Trp Ile Tyr Leu

145 150 155 160

Arg Pro Val Glu Thr Pro Thr Thr Val Phe Val Gln Ile Leu Arg Leu

165 170 175

Ile Lys Pro Met Lys Asp Gly Thr Arg Tyr Thr Gly Ile Arg Ser Leu

180 185 190

Lys Leu Asp Met Asn Pro Gly Thr Gly Ile Trp Gln Ser Ile Asp Val

195 200 205

Lys Thr Val Leu Gln Asn Trp Leu Lys Gln Pro Glu Ser Asn Leu Gly

210 215 220

Ile Glu Ile Lys Ala Leu Asp Glu Asn Gly His Asp Leu Ala Val Thr

225 230 235 240

Phe Pro Gly Pro Gly Glu Asp Gly Leu Asn Pro Phe Leu Glu Val Lys

245 250 255

Val Thr Asp Thr Pro Lys Arg Ser Arg Arg Asp Phe Gly Leu Asp Cys

260 265 270

Asp Glu His Ser Thr Glu Ser Arg Cys Cys Arg Tyr Pro Leu Thr Val

275 280 285

Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala Pro Lys Arg Tyr

290 295 300

Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu Phe Val Phe Leu Gln Lys

305 310 315 320

Tyr Pro His Thr His Leu Val His Gln Ala Asn Pro Arg Gly Ser Ala

325 330 335

Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile Asn Met Leu Tyr

340 345 350

Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly Lys Ile Pro Ala Met Val

355 360 365

Val Asp Arg Cys Gly Cys Ser

370 375

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Asp Phe Gly Leu Asp Cys Asp Glu Ser Thr Glu Ser Arg Cys Cys

1 5 10 15

Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile

20 25 30

Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu

35 40 45

Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala

50 55 60

Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser

65 70 75 80

Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly

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Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser

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Met Gln Lys Leu Gln Leu Cys Val Tyr Ile Tyr Leu Phe Met Leu Ile

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Val Ala Gly Pro Val Asp Leu Asn Glu Asn Ser Glu Gln Lys Glu Asn

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Val Glu Lys Glu Gly Leu Cys Asn Ala Cys Thr Trp Arg Gln Asn Thr

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Lys Ser Ser Arg Ile Glu Ala Ile Lys Ile Gln Ile Leu Ser Lys Leu

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Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Asp Ala Ile Arg Gln Leu

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Leu Pro Lys Ala Pro Pro Leu Arg Glu Leu Ile Asp Gln Tyr Asp Val

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Gln Arg Asp Asp Ser Ser Asp Gly Ser Leu Glu Asp Asp Asp Tyr His

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Ala Thr Thr Glu Thr Ile Ile Thr Met Pro Thr Glu Ser Asp Phe Leu

115 120 125

Met Gln Val Asp Gly Lys Pro Lys Cys Cys Phe Phe Lys Phe Ser Ser

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Lys Ile Gln Tyr Asn Lys Val Val Lys Ala Gln Leu Trp Ile Tyr Leu

145 150 155 160

Arg Pro Val Glu Thr Pro Thr Thr Val Phe Val Gln Ile Leu Arg Leu

165 170 175

Ile Lys Pro Met Lys Asp Gly Thr Arg Tyr Thr Gly Ile Arg Ser Leu

180 185 190

Lys Leu Asp Met Asn Pro Gly Thr Gly Ile Trp Gln Ser Ile Asp Val

195 200 205

Lys Thr Val Leu Gln Asn Trp Leu Lys Gln Pro Glu Ser Asn Leu Gly

210 215 220

Ile Glu Ile Lys Ala Leu Asp Glu Asn Gly His Asp Leu Ala Val Thr

225 230 235 240

Phe Pro Gly Pro Gly Glu Asp Gly Leu Asn Pro Phe Leu Glu Val Lys

245 250 255

Val Thr Asp Thr Pro Lys Arg Ser Arg Arg Asp Phe Gly Leu Asp Cys

260 265 270

Asp Glu His Ser Thr Glu Ser Arg Cys Cys Arg Tyr Pro Leu Thr Val

275 280 285

Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala Pro Lys Arg Tyr

290 295 300

Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu Phe Val Phe Leu Gln Lys

305 310 315 320

Tyr Pro His Thr His Leu Val His Gln Ala Asn Pro Arg Gly Ser Ala

325 330 335

Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile Asn Met Leu Tyr

340 345 350

Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly Lys Ile Pro Ala Met Val

355 360 365

Val Asp Arg Cys Gly Cys Ser

370 375

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Asp Phe Gly Leu Asp Cys Asp Glu Ser Thr Glu Ser Arg Cys Cys

1 5 10 15

Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile

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Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu

35 40 45

Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala

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Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser

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Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly

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Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser

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Met Met Gln Lys Leu Gln Met Tyr Val Tyr Ile Tyr Leu Phe Met Leu

1 5 10 15

Ile Ala Ala Gly Pro Val Asp Leu Asn Glu Gly Ser Glu Arg Glu Glu

20 25 30

Asn Val Glu Lys Glu Gly Leu Cys Asn Ala Cys Ala Trp Arg Gln Asn

35 40 45

Thr Arg Tyr Ser Arg Ile Glu Ala Ile Lys Ile Gln Ile Leu Ser Lys

50 55 60

Leu Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Asp Ala Ile Arg Gln

65 70 75 80

Leu Leu Pro Arg Ala Pro Pro Leu Arg Glu Leu Ile Asp Gln Tyr Asp

85 90 95

Val Gln Arg Asp Asp Ser Ser Asp Gly Ser Leu Glu Asp Asp Asp Tyr

100 105 110

His Ala Thr Thr Glu Thr Ile Ile Thr Met Pro Thr Glu Ser Asp Phe

115 120 125

Leu Met Gln Ala Asp Gly Lys Pro Lys Cys Cys Phe Phe Lys Phe Ser

130 135 140

Ser Lys Ile Gln Tyr Asn Lys Val Val Lys Ala Gln Leu Trp Ile Tyr

145 150 155 160

Leu Arg Pro Val Lys Thr Pro Thr Thr Val Phe Val Gln Ile Leu Arg

165 170 175

Leu Ile Lys Pro Met Lys Asp Gly Thr Arg Tyr Thr Gly Ile Arg Ser

180 185 190

Leu Lys Leu Asp Met Ser Pro Gly Thr Gly Ile Trp Gln Ser Ile Asp

195 200 205

Val Lys Thr Val Leu Gln Asn Trp Leu Lys Gln Pro Glu Ser Asn Leu

210 215 220

Gly Ile Glu Ile Lys Ala Leu Asp Glu Asn Gly His Asp Leu Ala Val

225 230 235 240

Thr Phe Pro Gly Pro Gly Glu Asp Gly Leu Asn Pro Phe Leu Glu Val

245 250 255

Lys Val Thr Asp Thr Pro Lys Arg Ser Arg Arg Asp Phe Gly Leu Asp

260 265 270

Cys Asp Glu His Ser Thr Glu Ser Arg Cys Cys Arg Tyr Pro Leu Thr

275 280 285

Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala Pro Lys Arg

290 295 300

Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu Phe Val Phe Leu Gln

305 310 315 320

Lys Tyr Pro His Thr His Leu Val His Gln Ala Asn Pro Arg Gly Ser

325 330 335

Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile Asn Met Leu

340 345 350

Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly Lys Ile Pro Ala Met

355 360 365

Val Val Asp Arg Cys Gly Cys Ser

370 375

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<212> PRT

<213> Mus musculus

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Asp Phe Gly Leu Asp Cys Asp Glu Ser Thr Glu Ser Arg Cys Cys

1 5 10 15

Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile

20 25 30

Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Glu Cys Glu

35 40 45

Phe Val Phe Leu Gln Lys Tyr Pro His Thr His Leu Val His Gln Ala

50 55 60

Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser

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Pro Ile Asn Met Leu Tyr Phe Asn Gly Lys Glu Gln Ile Ile Tyr Gly

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Lys Ile Pro Ala Met Val Val Asp Arg Cys Gly Cys Ser

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<210> 7

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<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

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Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 8

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 8

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Cys

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 9

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 9

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 10

<211> 107

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 10

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 11

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 11

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Arg Gly Gly Pro Lys Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 12

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 12

Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Gly Ser

1 5 10 15

Leu Thr Val Thr Cys Arg Val Ser Gly Ile Asp Leu Ser Ser Tyr Asp

20 25 30

Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

35 40 45

Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Met Thr

65 70 75 80

Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Val

85 90 95

Pro Ala Tyr Ser Thr Gly Gly Asp Leu Trp Gly Pro Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 13

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 13

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 14

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 14

Ala Gln Val Leu Thr Gln Thr Ala Ser Ser Val Ser Ala Ala Val Gly

1 5 10 15

Gly Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Ser Val Tyr Asp Asn

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe

50 55 60

Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Asp Leu

65 70 75 80

Glu Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Leu Tyr Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 15

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 15

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Gln Ser Ser Gln Ser Val Tyr Asp Asn

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Leu Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 16

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 16

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 17

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 17

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 18

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 18

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 19

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 19

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 20

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 20

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 21

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 21

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 22

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 22

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 23

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 23

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser Thr Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 24

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 24

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 25

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 25

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 26

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 26

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 27

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 27

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 28

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 28

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 29

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 29

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 30

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 30

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala His Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 31

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 31

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Gln Ser Ser Gln Ser Val Tyr His Asn

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Leu Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 32

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 32

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Thr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 33

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 33

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Thr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Met Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 34

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 34

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Tyr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 35

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 35

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Thr Ser Ser Gln Ser Val Tyr His Asn

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Tyr Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Arg Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 36

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 36

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Gln Ser Ser Gln Ser Val Tyr His Asn

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Phe Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Leu Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 37

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 37

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Gln Thr Ser Gln Ser Val Tyr His Asp

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Arg Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 38

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 38

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Thr Ser Ser Gln Ser Val Tyr His Asp

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Tyr Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Leu Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 39

<211> 348

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 39

cagtcgctgg aggagtccgg gggtcgcctg gtaacgcctg gaggatccct gacagtcacc 60

tgcagagtct ctggaatcga cctcagtagt tacgacatca gctgggtccg ccaggctcca 120

gggaaggggc tggagtggat cggaatcatt agttatgctg gtagcacata ctacgcgagc 180

tgggcaaaag gccgattcac catctccaaa acctcgacca cggtggatct gaaaatgacc 240

agtctgacaa ccgaggacac ggccacctat ttctgtgcca gaggtgtgcc tgcttatagt 300

actggtggtg acttgtgggg cccaggcacc ctggtcaccg tctcctcc 348

<210> 40

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 40

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc agctacgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggagcac atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactcta ccggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 41

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 41

gcgcaagtgc tgacccagac tgcatcatcc gtgtctgcag ctgtgggagg cacagtcacc 60

120

aaaccagggc agcctcccaa gctcctgatc tattgggcat ccactctggc atctggggtc 180

ccatcgcggt tcaaaggcag tggatctggg acacagttca ctctcaccat cagcgacctg 240

gagtgtgacg atgctgccac ttactactgt gcaggcggtt atggtggtgg tctgtatgct 300

ttcggcggag ggaccaaggt ggagatcaaa 330

<210> 42

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 42

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgcc agagcagcca gtccgtgtac gacaacaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc cagcggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gctgtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 43

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 43

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacaccgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggagcac atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 44

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 44

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacaccgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggagcac atactatatg 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 45

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 45

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cactacgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggagcac atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 46

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 46

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgca ccagcagcca gtccgtgtac cacaacaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc ctacggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gcggtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 47

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 47

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgcc agagcagcca gtccgtgtac cacaacaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc cttcggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gctgtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 48

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 48

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgcc agaccagcca gtccgtgtac cacgacaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc cagcggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gcggtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 49

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 49

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgca ccagcagcca gtccgtgtac cacgacaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc ctacggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gctgtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 50

<211> 987

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 50

gcttccacca agggcccatc ggtcttcccc ctggcaccct cctccaagtc caccctctggg 60

ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 120

tggaactcag gcgccctgac ctccggcgtg cacaccttcc cggctgtcct acagtcctca 180

ggactctact ccctctcctc cgtggtgacc gtgccctcct cgtccttgggg cacccagacc 240

tacatctgca acgtgaatca caagccctcc aacaccaagg tggacaagaa agttgagccc 300

aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga 360

ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 420

gaggtcacat gcgtggtggt ggacgtgtcc cacgaagacc ctgaggtcaa gttcaactgg 480

tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 540

tccacgtacc gtgtggtctc cgtcctcacc gtcctgcacc aggactggct gaatggcaag 600

gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc 660

aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggaggaa 720

atgaccaaga accaggtctc cctgacctgc ctggtcaaag gcttctatcc ctccgacatc 780

gccgtggagt gggagtccaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 840

ctggactccg acggctcctt cttcctctac tccaagctca ccgtggacaa gtccaggtgg 900

cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 960

cagaagtcgc tctccctgtc tccgtag 987

<210> 51

<211> 984

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 51

gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag caccctctggg 60

ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 120

tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 180

ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 240

tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 300

aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaact ccggggggga 360

ccgaaagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 420

gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 480

tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 540

agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 600

gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc 660

aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggatgag 720

ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 780

gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 840

ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 900

cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 960

cagaagagcc tctccctgtc tccg 984

<210> 52

<211> 987

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 52

gcttccacca agggcccatc ggtcttcccc ctggcaccct cctccaagtc gaccctctggg 60

ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 120

tggaactcag gcgccctgac cagtggcgtg cacaccttcc cggctgtcct acagtcctca 180

ggactctact ccctcagtag tgtggtgacc gtgccctcca gtagtttgggg cacccagacc 240

tacatctgca acgtgaatca caagcccagt aacaccaagg tggacaagaa agttgagccc 300

aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgagct cctgggggga 360

ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 420

gaggtcacat gcgtggtggt ggacgtgagt cacgaagacc ctgaggtcaa gttcaactgg 480

tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 540

agtacgtacc gtgtggtcag tgtcctcacc gtcctgcacc aggactggct gaatggcaag 600

gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc 660

aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggaggag 720

atgaccaaga accaggtcag tctgacctgc ctggtcaaag gcttctatcc cagtgacatc 780

gccgtggagt gggagagtaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 840

ctggactccg acggctcctt cttcctctac agtaagctca ccgtggacaa gagtaggtgg 900

cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 960

cagaagagtc tctccctgtc tccgtga 987

<210> 53

<211> 324

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 53

cgtacggtgg ctgcaccatc tgtcttcatc ttcccgccat ctgatgagca gttgaaatct 60

ggaactgcct ctgttgtgtg cctgctgaat aacttctatc ccagagaggc caaagtacag 120

tggaaggtgg ataacgccct ccaatcgggt aactcccagg agagtgtcac agagcaggac 180

tccaaggact gcacctactc cctctcctcc accctgacgc tgtccaaagc agactacgag 240

aaacacaaag tctacgcctg cgaagtcacc catcagggcc tgtcctcgcc cgtcacaaag 300

tccttcaaca ggggagagtg ttga 324

<210> 54

<211> 324

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 54

cgtacggtgg ctgcaccatc tgtcttcatc ttcccgccat ctgatgagca gttgaaatct 60

ggtaccgcct ctgttgtgtg cctgctgaat aacttctatc ccagagaggc caaagtacag 120

tggaaggtgg ataacgccct ccaatcgggt aactcccagg agagtgtcac agagcaggac 180

agtaaggaca gtacctacag tctcagtagt accctgacgc tgtccaaagc agactacgag 240

aaacacaaag tctacgcctg cgaagtcacc catcagggcc tgtcctcgcc cgtcacaaag 300

tccttcaaca ggggagagtg ttga 324

<210> 55

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 55

Ser Tyr Asp Ile Ser

15

<210> 56

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 56

His Tyr Asp Ile Ser

15

<210> 57

<211> 5

<212> PRT

<213> Artificial Sequence

<220>

<223> An artificially synthesized sequence

<400> 57

His Thr Asp Ile Ser

15

<210> 58

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 58

Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 59

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 59

Ile Ile Ser His Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 60

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 60

Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Met Ser Trp Ala Lys Gly

1 5 10 15

<210> 61

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 61

Gly Val Pro Ala Tyr Ser Thr Gly Gly Asp Leu

1 5 10

<210> 62

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 62

Gly Val Pro Ala His Ser Thr Gly Gly Asp Leu

1 5 10

<210> 63

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 63

Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu

1 5 10

<210> 64

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 64

Gly Val Pro Ala His Ser His Gly Gly Asp Leu

1 5 10

<210> 65

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 65

Gln Ser Ser Gln Ser Val Tyr Asp Asn Asn Trp Leu Ser

1 5 10

<210> 66

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 66

Gln Ser Ser Gln Ser Val Tyr His Asn Asn Trp Leu Ser

1 5 10

<210> 67

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 67

Thr Ser Ser Gln Ser Val Tyr His Asn Asn Trp Leu Ser

1 5 10

<210> 68

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 68

Gln Thr Ser Gln Ser Val Tyr His Asp Asn Trp Leu Ser

1 5 10

<210> 69

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 69

Thr Ser Ser Gln Ser Val Tyr His Asp Asn Trp Leu Ser

1 5 10

<210> 70

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 70

Trp Ala Ser Thr Leu Ala Ser

15

<210> 71

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 71

Trp Ala Ser Thr Leu Ala Tyr

15

<210> 72

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 72

Trp Ala Ser Thr Leu Ala Phe

15

<210> 73

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 73

Ala Gly Gly Tyr Gly Gly Gly Leu Tyr Ala

1 5 10

<210> 74

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 74

Ala Gly Gly Tyr Gly Gly Gly Arg Tyr Ala

1 5 10

<210> 75

<211> 242

<212> PRT

<213> Homo sapiens

<400> 75

Glu Asn Ser Glu Gln Lys Glu Asn Val Glu Lys Glu Gly Leu Cys Asn

1 5 10 15

Ala Cys Thr Trp Arg Gln Asn Thr Lys Ser Ser Arg Ile Glu Ala Ile

20 25 30

Lys Ile Gln Ile Leu Ser Lys Leu Arg Leu Glu Thr Ala Pro Asn Ile

35 40 45

Ser Lys Asp Val Ile Arg Gln Leu Leu Pro Lys Ala Pro Pro Leu Arg

50 55 60

Glu Leu Ile Asp Gln Tyr Asp Val Gln Arg Asp Asp Ser Ser Asp Gly

65 70 75 80

Ser Leu Glu Asp Asp Asp Tyr His Ala Thr Thr Glu Thr Ile Ile Thr

85 90 95

Met Pro Thr Glu Ser Asp Phe Leu Met Gln Val Asp Gly Lys Pro Lys

100 105 110

Cys Cys Phe Phe Lys Phe Ser Ser Lys Ile Gln Tyr Asn Lys Val Val

115 120 125

Lys Ala Gln Leu Trp Ile Tyr Leu Arg Pro Val Glu Thr Pro Thr Thr

130 135 140

Val Phe Val Gln Ile Leu Arg Leu Ile Lys Pro Met Lys Asp Gly Thr

145 150 155 160

Arg Tyr Thr Gly Ile Arg Ser Leu Lys Leu Asp Met Asn Pro Gly Thr

165 170 175

Gly Ile Trp Gln Ser Ile Asp Val Lys Thr Val Leu Gln Asn Trp Leu

180 185 190

Lys Gln Pro Glu Ser Asn Leu Gly Ile Glu Ile Lys Ala Leu Asp Glu

195 200 205

Asn Gly His Asp Leu Ala Val Thr Phe Pro Gly Pro Gly Glu Asp Gly

210 215 220

Leu Asn Pro Phe Leu Glu Val Lys Val Thr Asp Thr Pro Lys Arg Ser

225 230 235 240

Arg Arg

<210> 76

<211> 242

<212> PRT

<213> Macaca fascicularis

<400> 76

Glu Asn Ser Glu Gln Lys Glu Asn Val Glu Lys Glu Gly Leu Cys Asn

1 5 10 15

Ala Cys Thr Trp Arg Gln Asn Thr Lys Ser Ser Arg Ile Glu Ala Ile

20 25 30

Lys Ile Gln Ile Leu Ser Lys Leu Arg Leu Glu Thr Ala Pro Asn Ile

35 40 45

Ser Lys Asp Ala Ile Arg Gln Leu Leu Pro Lys Ala Pro Pro Leu Arg

50 55 60

Glu Leu Ile Asp Gln Tyr Asp Val Gln Arg Asp Asp Ser Ser Asp Gly

65 70 75 80

Ser Leu Glu Asp Asp Asp Tyr His Ala Thr Thr Glu Thr Ile Ile Thr

85 90 95

Met Pro Thr Glu Ser Asp Phe Leu Met Gln Val Asp Gly Lys Pro Lys

100 105 110

Cys Cys Phe Phe Lys Phe Ser Ser Lys Ile Gln Tyr Asn Lys Val Val

115 120 125

Lys Ala Gln Leu Trp Ile Tyr Leu Arg Pro Val Glu Thr Pro Thr Thr

130 135 140

Val Phe Val Gln Ile Leu Arg Leu Ile Lys Pro Met Lys Asp Gly Thr

145 150 155 160

Arg Tyr Thr Gly Ile Arg Ser Leu Lys Leu Asp Met Asn Pro Gly Thr

165 170 175

Gly Ile Trp Gln Ser Ile Asp Val Lys Thr Val Leu Gln Asn Trp Leu

180 185 190

Lys Gln Pro Glu Ser Asn Leu Gly Ile Glu Ile Lys Ala Leu Asp Glu

195 200 205

Asn Gly His Asp Leu Ala Val Thr Phe Pro Gly Pro Gly Glu Asp Gly

210 215 220

Leu Asn Pro Phe Leu Glu Val Lys Val Thr Asp Thr Pro Lys Arg Ser

225 230 235 240

Arg Arg

<210> 77

<211> 242

<212> PRT

<213> Mus musculus

<400> 77

Glu Gly Ser Glu Arg Glu Glu Asn Val Glu Lys Glu Gly Leu Cys Asn

1 5 10 15

Ala Cys Ala Trp Arg Gln Asn Thr Arg Tyr Ser Arg Ile Glu Ala Ile

20 25 30

Lys Ile Gln Ile Leu Ser Lys Leu Arg Leu Glu Thr Ala Pro Asn Ile

35 40 45

Ser Lys Asp Ala Ile Arg Gln Leu Leu Pro Arg Ala Pro Pro Leu Arg

50 55 60

Glu Leu Ile Asp Gln Tyr Asp Val Gln Arg Asp Asp Ser Ser Asp Gly

65 70 75 80

Ser Leu Glu Asp Asp Asp Tyr His Ala Thr Thr Glu Thr Ile Ile Thr

85 90 95

Met Pro Thr Glu Ser Asp Phe Leu Met Gln Ala Asp Gly Lys Pro Lys

100 105 110

Cys Cys Phe Phe Lys Phe Ser Ser Lys Ile Gln Tyr Asn Lys Val Val

115 120 125

Lys Ala Gln Leu Trp Ile Tyr Leu Arg Pro Val Lys Thr Pro Thr Thr

130 135 140

Val Phe Val Gln Ile Leu Arg Leu Ile Lys Pro Met Lys Asp Gly Thr

145 150 155 160

Arg Tyr Thr Gly Ile Arg Ser Leu Lys Leu Asp Met Ser Pro Gly Thr

165 170 175

Gly Ile Trp Gln Ser Ile Asp Val Lys Thr Val Leu Gln Asn Trp Leu

180 185 190

Lys Gln Pro Glu Ser Asn Leu Gly Ile Glu Ile Lys Ala Leu Asp Glu

195 200 205

Asn Gly His Asp Leu Ala Val Thr Phe Pro Gly Pro Gly Glu Asp Gly

210 215 220

Leu Asn Pro Phe Leu Glu Val Lys Val Thr Asp Thr Pro Lys Arg Ser

225 230 235 240

Arg Arg

<210> 78

<211> 266

<212> PRT

<213> Homo sapiens

<400> 78

Met Gln Lys Leu Gln Leu Cys Val Tyr Ile Tyr Leu Phe Met Leu Ile

1 5 10 15

Val Ala Gly Pro Val Asp Leu Asn Glu Asn Ser Glu Gln Lys Glu Asn

20 25 30

Val Glu Lys Glu Gly Leu Cys Asn Ala Cys Thr Trp Arg Gln Asn Thr

35 40 45

Lys Ser Ser Arg Ile Glu Ala Ile Lys Ile Gln Ile Leu Ser Lys Leu

50 55 60

Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Asp Val Ile Arg Gln Leu

65 70 75 80

Leu Pro Lys Ala Pro Pro Leu Arg Glu Leu Ile Asp Gln Tyr Asp Val

85 90 95

Gln Arg Asp Asp Ser Ser Asp Gly Ser Leu Glu Asp Asp Asp Tyr His

100 105 110

Ala Thr Thr Glu Thr Ile Ile Thr Met Pro Thr Glu Ser Asp Phe Leu

115 120 125

Met Gln Val Asp Gly Lys Pro Lys Cys Cys Phe Phe Lys Phe Ser Ser

130 135 140

Lys Ile Gln Tyr Asn Lys Val Val Lys Ala Gln Leu Trp Ile Tyr Leu

145 150 155 160

Arg Pro Val Glu Thr Pro Thr Thr Val Phe Val Gln Ile Leu Arg Leu

165 170 175

Ile Lys Pro Met Lys Asp Gly Thr Arg Tyr Thr Gly Ile Arg Ser Leu

180 185 190

Lys Leu Asp Met Asn Pro Gly Thr Gly Ile Trp Gln Ser Ile Asp Val

195 200 205

Lys Thr Val Leu Gln Asn Trp Leu Lys Gln Pro Glu Ser Asn Leu Gly

210 215 220

Ile Glu Ile Lys Ala Leu Asp Glu Asn Gly His Asp Leu Ala Val Thr

225 230 235 240

Phe Pro Gly Pro Gly Glu Asp Gly Leu Asn Pro Phe Leu Glu Val Lys

245 250 255

Val Thr Asp Thr Pro Lys Arg Ser Arg Arg

260 265

<210> 79

<211> 266

<212> PRT

<213> Macaca fascicularis

<400> 79

Met Gln Lys Leu Gln Leu Cys Val Tyr Ile Tyr Leu Phe Met Leu Ile

1 5 10 15

Val Ala Gly Pro Val Asp Leu Asn Glu Asn Ser Glu Gln Lys Glu Asn

20 25 30

Val Glu Lys Glu Gly Leu Cys Asn Ala Cys Thr Trp Arg Gln Asn Thr

35 40 45

Lys Ser Ser Arg Ile Glu Ala Ile Lys Ile Gln Ile Leu Ser Lys Leu

50 55 60

Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Asp Ala Ile Arg Gln Leu

65 70 75 80

Leu Pro Lys Ala Pro Pro Leu Arg Glu Leu Ile Asp Gln Tyr Asp Val

85 90 95

Gln Arg Asp Asp Ser Ser Asp Gly Ser Leu Glu Asp Asp Asp Tyr His

100 105 110

Ala Thr Thr Glu Thr Ile Ile Thr Met Pro Thr Glu Ser Asp Phe Leu

115 120 125

Met Gln Val Asp Gly Lys Pro Lys Cys Cys Phe Phe Lys Phe Ser Ser

130 135 140

Lys Ile Gln Tyr Asn Lys Val Val Lys Ala Gln Leu Trp Ile Tyr Leu

145 150 155 160

Arg Pro Val Glu Thr Pro Thr Thr Val Phe Val Gln Ile Leu Arg Leu

165 170 175

Ile Lys Pro Met Lys Asp Gly Thr Arg Tyr Thr Gly Ile Arg Ser Leu

180 185 190

Lys Leu Asp Met Asn Pro Gly Thr Gly Ile Trp Gln Ser Ile Asp Val

195 200 205

Lys Thr Val Leu Gln Asn Trp Leu Lys Gln Pro Glu Ser Asn Leu Gly

210 215 220

Ile Glu Ile Lys Ala Leu Asp Glu Asn Gly His Asp Leu Ala Val Thr

225 230 235 240

Phe Pro Gly Pro Gly Glu Asp Gly Leu Asn Pro Phe Leu Glu Val Lys

245 250 255

Val Thr Asp Thr Pro Lys Arg Ser Arg Arg

260 265

<210> 80

<211> 267

<212> PRT

<213> Mus musculus

<400> 80

Met Met Gln Lys Leu Gln Met Tyr Val Tyr Ile Tyr Leu Phe Met Leu

1 5 10 15

Ile Ala Ala Gly Pro Val Asp Leu Asn Glu Gly Ser Glu Arg Glu Glu

20 25 30

Asn Val Glu Lys Glu Gly Leu Cys Asn Ala Cys Ala Trp Arg Gln Asn

35 40 45

Thr Arg Tyr Ser Arg Ile Glu Ala Ile Lys Ile Gln Ile Leu Ser Lys

50 55 60

Leu Arg Leu Glu Thr Ala Pro Asn Ile Ser Lys Asp Ala Ile Arg Gln

65 70 75 80

Leu Leu Pro Arg Ala Pro Pro Leu Arg Glu Leu Ile Asp Gln Tyr Asp

85 90 95

Val Gln Arg Asp Asp Ser Ser Asp Gly Ser Leu Glu Asp Asp Asp Tyr

100 105 110

His Ala Thr Thr Glu Thr Ile Ile Thr Met Pro Thr Glu Ser Asp Phe

115 120 125

Leu Met Gln Ala Asp Gly Lys Pro Lys Cys Cys Phe Phe Lys Phe Ser

130 135 140

Ser Lys Ile Gln Tyr Asn Lys Val Val Lys Ala Gln Leu Trp Ile Tyr

145 150 155 160

Leu Arg Pro Val Lys Thr Pro Thr Thr Val Phe Val Gln Ile Leu Arg

165 170 175

Leu Ile Lys Pro Met Lys Asp Gly Thr Arg Tyr Thr Gly Ile Arg Ser

180 185 190

Leu Lys Leu Asp Met Ser Pro Gly Thr Gly Ile Trp Gln Ser Ile Asp

195 200 205

Val Lys Thr Val Leu Gln Asn Trp Leu Lys Gln Pro Glu Ser Asn Leu

210 215 220

Gly Ile Glu Ile Lys Ala Leu Asp Glu Asn Gly His Asp Leu Ala Val

225 230 235 240

Thr Phe Pro Gly Pro Gly Glu Asp Gly Leu Asn Pro Phe Leu Glu Val

245 250 255

Lys Val Thr Asp Thr Pro Lys Arg Ser Arg Arg

260 265

<210> 81

<211> 407

<212> PRT

<213> Homo sapiens

<400> 81

Met Val Leu Ala Ala Pro Leu Leu Leu Gly Phe Leu Leu Leu Ala Leu

1 5 10 15

Glu Leu Arg Pro Arg Gly Glu Ala Ala Glu Gly Pro Ala Ala Ala Ala

20 25 30

Ala Ala Ala Ala Ala Ala Ala Ala Ala Gly Val Gly Gly Glu Arg Ser

35 40 45

Ser Arg Pro Ala Pro Ser Val Ala Pro Glu Pro Asp Gly Cys Pro Val

50 55 60

Cys Val Trp Arg Gln His Ser Arg Glu Leu Arg Leu Glu Ser Ile Lys

65 70 75 80

Ser Gln Ile Leu Ser Lys Leu Arg Leu Lys Glu Ala Pro Asn Ile Ser

85 90 95

Arg Glu Val Val Lys Gln Leu Leu Pro Lys Ala Pro Pro Leu Gln Gln

100 105 110

Ile Leu Asp Leu His Asp Phe Gln Gly Asp Ala Leu Gln Pro Glu Asp

115 120 125

Phe Leu Glu Glu Asp Glu Tyr His Ala Thr Thr Glu Thr Val Ile Ser

130 135 140

Met Ala Gln Glu Thr Asp Pro Ala Val Gln Thr Asp Gly Ser Pro Leu

145 150 155 160

Cys Cys His Phe His Phe Ser Pro Lys Val Met Phe Thr Lys Val Leu

165 170 175

Lys Ala Gln Leu Trp Val Tyr Leu Arg Pro Val Pro Arg Pro Ala Thr

180 185 190

Val Tyr Leu Gln Ile Leu Arg Leu Lys Pro Leu Thr Gly Glu Gly Thr

195 200 205

Ala Gly Gly Gly Gly Gly Gly Arg Arg His Ile Arg Ile Arg Ser Leu

210 215 220

Lys Ile Glu Leu His Ser Arg Ser Gly His Trp Gln Ser Ile Asp Phe

225 230 235 240

Lys Gln Val Leu His Ser Trp Phe Arg Gln Pro Gln Ser Asn Trp Gly

245 250 255

Ile Glu Ile Asn Ala Phe Asp Pro Ser Gly Thr Asp Leu Ala Val Thr

260 265 270

Ser Leu Gly Pro Gly Ala Glu Gly Leu His Pro Phe Met Glu Leu Arg

275 280 285

Val Leu Glu Asn Thr Lys Arg Ser Arg Arg Asn Leu Gly Leu Asp Cys

290 295 300

Asp Glu His Ser Ser Glu Ser Arg Cys Cys Arg Tyr Pro Leu Thr Val

305 310 315 320

Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala Pro Lys Arg Tyr

325 330 335

Lys Ala Asn Tyr Cys Ser Gly Gln Cys Glu Tyr Met Phe Met Gln Lys

340 345 350

Tyr Pro His Thr His Leu Val Gln Gln Ala Asn Pro Arg Gly Ser Ala

355 360 365

Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile Asn Met Leu Tyr

370 375 380

Phe Asn Asp Lys Gln Gln Ile Ile Tyr Gly Lys Ile Pro Gly Met Val

385 390 395 400

Val Asp Arg Cys Gly Cys Ser

405

<210> 82

<211> 109

<212> PRT

<213> Homo sapiens

<400> 82

Asn Leu Gly Leu Asp Cys Asp Glu Ser Ser Glu Ser Arg Cys Cys

1 5 10 15

Arg Tyr Pro Leu Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile

20 25 30

Ile Ala Pro Lys Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Gln Cys Glu

35 40 45

Tyr Met Phe Met Gln Lys Tyr Pro His Thr His Leu Val Gln Gln Ala

50 55 60

Asn Pro Arg Gly Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser

65 70 75 80

Pro Ile Asn Met Leu Tyr Phe Asn Asp Lys Gln Gln Ile Ile Tyr Gly

85 90 95

Lys Ile Pro Gly Met Val Val Asp Arg Cys Gly Cys Ser

100 105

<210> 83

<211> 274

<212> PRT

<213> Homo sapiens

<400> 83

Ala Glu Gly Pro Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala

1 5 10 15

Ala Gly Val Gly Gly Glu Arg Ser Ser Arg Pro Ala Pro Ser Val Ala

20 25 30

Pro Glu Pro Asp Gly Cys Pro Val Cys Val Trp Arg Gln His Ser Arg

35 40 45

Glu Leu Arg Leu Glu Ser Ile Lys Ser Gln Ile Leu Ser Lys Leu Arg

50 55 60

Leu Lys Glu Ala Pro Asn Ile Ser Arg Glu Val Val Lys Gln Leu Leu

65 70 75 80

Pro Lys Ala Pro Pro Leu Gln Gln Ile Leu Asp Leu His Asp Phe Gln

85 90 95

Gly Asp Ala Leu Gln Pro Glu Asp Phe Leu Glu Glu Asp Glu Tyr His

100 105 110

Ala Thr Thr Glu Thr Val Ile Ser Met Ala Gln Glu Thr Asp Pro Ala

115 120 125

Val Gln Thr Asp Gly Ser Pro Leu Cys Cys His Phe His Phe Ser Pro

130 135 140

Lys Val Met Phe Thr Lys Val Leu Lys Ala Gln Leu Trp Val Tyr Leu

145 150 155 160

Arg Pro Val Pro Arg Pro Ala Thr Val Tyr Leu Gln Ile Leu Arg Leu

165 170 175

Lys Pro Leu Thr Gly Glu Gly Thr Ala Gly Gly Gly Gly Gly Gly Arg

180 185 190

Arg His Ile Arg Ile Arg Ser Leu Lys Ile Glu Leu His Ser Arg Ser

195 200 205

Gly His Trp Gln Ser Ile Asp Phe Lys Gln Val Leu His Ser Trp Phe

210 215 220

Arg Gln Pro Gln Ser Asn Trp Gly Ile Glu Ile Asn Ala Phe Asp Pro

225 230 235 240

Ser Gly Thr Asp Leu Ala Val Thr Ser Leu Gly Pro Gly Ala Glu Gly

245 250 255

Leu His Pro Phe Met Glu Leu Arg Val Leu Glu Asn Thr Lys Arg Ser

260 265 270

Arg Arg

<210> 84

<211> 298

<212> PRT

<213> Homo sapiens

<400> 84

Met Val Leu Ala Ala Pro Leu Leu Leu Gly Phe Leu Leu Leu Ala Leu

1 5 10 15

Glu Leu Arg Pro Arg Gly Glu Ala Ala Glu Gly Pro Ala Ala Ala Ala

20 25 30

Ala Ala Ala Ala Ala Ala Ala Ala Ala Gly Val Gly Gly Glu Arg Ser

35 40 45

Ser Arg Pro Ala Pro Ser Val Ala Pro Glu Pro Asp Gly Cys Pro Val

50 55 60

Cys Val Trp Arg Gln His Ser Arg Glu Leu Arg Leu Glu Ser Ile Lys

65 70 75 80

Ser Gln Ile Leu Ser Lys Leu Arg Leu Lys Glu Ala Pro Asn Ile Ser

85 90 95

Arg Glu Val Val Lys Gln Leu Leu Pro Lys Ala Pro Pro Leu Gln Gln

100 105 110

Ile Leu Asp Leu His Asp Phe Gln Gly Asp Ala Leu Gln Pro Glu Asp

115 120 125

Phe Leu Glu Glu Asp Glu Tyr His Ala Thr Thr Glu Thr Val Ile Ser

130 135 140

Met Ala Gln Glu Thr Asp Pro Ala Val Gln Thr Asp Gly Ser Pro Leu

145 150 155 160

Cys Cys His Phe His Phe Ser Pro Lys Val Met Phe Thr Lys Val Leu

165 170 175

Lys Ala Gln Leu Trp Val Tyr Leu Arg Pro Val Pro Arg Pro Ala Thr

180 185 190

Val Tyr Leu Gln Ile Leu Arg Leu Lys Pro Leu Thr Gly Glu Gly Thr

195 200 205

Ala Gly Gly Gly Gly Gly Gly Arg Arg His Ile Arg Ile Arg Ser Leu

210 215 220

Lys Ile Glu Leu His Ser Arg Ser Gly His Trp Gln Ser Ile Asp Phe

225 230 235 240

Lys Gln Val Leu His Ser Trp Phe Arg Gln Pro Gln Ser Asn Trp Gly

245 250 255

Ile Glu Ile Asn Ala Phe Asp Pro Ser Gly Thr Asp Leu Ala Val Thr

260 265 270

Ser Leu Gly Pro Gly Ala Glu Gly Leu His Pro Phe Met Glu Leu Arg

275 280 285

Val Leu Glu Asn Thr Lys Arg Ser Arg Arg

290 295

<210> 85

<211> 409

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 85

Met Lys Trp Val Thr Phe Leu Leu Leu Leu Phe Ile Ser Gly Ser Ala

1 5 10 15

Phe Ser Asp Tyr Lys Asp Asp Asp Asp Lys Ala Glu Gly Pro Ala Ala

20 25 30

Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Gly Val Gly Gly Glu

35 40 45

Arg Ser Ser Arg Pro Ala Pro Ser Val Ala Pro Glu Pro Asp Gly Cys

50 55 60

Pro Val Cys Val Trp Arg Gln His Ser Arg Glu Leu Arg Leu Glu Ser

65 70 75 80

Ile Lys Ser Gln Ile Leu Ser Lys Leu Arg Leu Lys Glu Ala Pro Asn

85 90 95

Ile Ser Arg Glu Val Val Lys Gln Leu Leu Pro Lys Ala Pro Pro Leu

100 105 110

Gln Gln Ile Leu Asp Leu His Asp Phe Gln Gly Asp Ala Leu Gln Pro

115 120 125

Glu Asp Phe Leu Glu Glu Asp Glu Tyr His Ala Thr Thr Glu Thr Val

130 135 140

Ile Ser Met Ala Gln Glu Thr Asp Pro Ala Val Gln Thr Asp Gly Ser

145 150 155 160

Pro Leu Cys Cys His Phe His Phe Ser Pro Lys Val Met Phe Thr Lys

165 170 175

Val Leu Lys Ala Gln Leu Trp Val Tyr Leu Arg Pro Val Pro Arg Pro

180 185 190

Ala Thr Val Tyr Leu Gln Ile Leu Arg Leu Lys Pro Leu Thr Gly Glu

195 200 205

Gly Thr Ala Gly Gly Gly Gly Gly Gly Arg Arg His Ile Arg Ile Arg

210 215 220

Ser Leu Lys Ile Glu Leu His Ser Arg Ser Gly His Trp Gln Ser Ile

225 230 235 240

Asp Phe Lys Gln Val Leu His Ser Trp Phe Arg Gln Pro Gln Ser Asn

245 250 255

Trp Gly Ile Glu Ile Asn Ala Phe Asp Pro Ser Gly Thr Asp Leu Ala

260 265 270

Val Thr Ser Leu Gly Pro Gly Ala Glu Gly Leu His Pro Phe Met Glu

275 280 285

Leu Arg Val Leu Glu Asn Thr Lys Arg Ser Arg Arg Asn Leu Gly Leu

290 295 300

Asp Cys Asp Glu His Ser Ser Glu Ser Arg Cys Cys Arg Tyr Pro Leu

305 310 315 320

Thr Val Asp Phe Glu Ala Phe Gly Trp Asp Trp Ile Ile Ala Pro Lys

325 330 335

Arg Tyr Lys Ala Asn Tyr Cys Ser Gly Gln Cys Glu Tyr Met Phe Met

340 345 350

Gln Lys Tyr Pro His Thr His Leu Val Gln Gln Ala Asn Pro Arg Gly

355 360 365

Ser Ala Gly Pro Cys Cys Thr Pro Thr Lys Met Ser Pro Ile Asn Met

370 375 380

Leu Tyr Phe Asn Asp Lys Gln Gln Ile Ile Tyr Gly Lys Ile Pro Gly

385 390 395 400

Met Val Val Asp Arg Cys Gly Cys Ser

405

<210> 86

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 86

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Asp

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 87

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 87

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Asp

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Met Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Lys Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 88

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 88

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Thr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Lys Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Lys Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 89

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 89

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Thr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Lys Lys Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 90

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 90

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Thr

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Ser Lys Tyr Lys Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 91

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 91

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser His Asp

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ile Ile Ser Tyr Ala Gly Lys Lys Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 92

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 92

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Ile Asp Leu Ser His Asp

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 93

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 93

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Lys Val Ser Gly Ile Asp Leu Ser His Asp

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 94

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 94

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Lys Val Ser Gly Ile Asp Leu Ser His Glu

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ile Ile Ser Tyr Ala Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 95

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 95

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Ile Asp Leu Ser His Glu

20 25 30

Asp Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ile Ile Ser Tyr Ala Gly Ser Lys Tyr Tyr Ala Ser Trp Ala Lys

50 55 60

Gly Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu

65 70 75 80

Thr Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala

85 90 95

Arg Gly Val Pro Ala Tyr Ser His Gly Gly Asp Leu Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 96

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 96

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Thr Ser Ser Gln Ser Val Tyr His Ala

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Tyr Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Arg Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 97

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 97

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Thr Thr Ser Gln Ser Val Tyr His Glu

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Tyr Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Arg Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 98

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 98

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Thr Ser Ser Gln Ser Val Phe His Glu

20 25 30

Asn Trp Leu Ser Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Trp Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Arg Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 99

<211> 110

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 99

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Thr Thr Ser Gln Ser Val Tyr His Glu

20 25 30

Asn Trp Leu Ser Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Trp Ala Ser Thr Leu Ala Tyr Gly Val Pro Ser Arg Phe

50 55 60

Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu

65 70 75 80

Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys Ala Gly Gly Tyr Gly Gly

85 90 95

Gly Arg Tyr Ala Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 100

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 100

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacgacgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggagcac atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 101

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 101

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacgacgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggagcat gtactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga taagtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 102

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 102

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacaccgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggaagac atactatgcc 180

agctgggcta aaaaaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 103

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 103

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacaccgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggaagaa atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 104

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 104

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacaccgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggagcaa atacaaggcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 105

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 105

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcacag tgagtgggat cgacctgagc cacgacgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtcggcatc attagctacg ccgggaagaa atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 106

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 106

caggtgcagc tggtcgaatc tggaggagga ctggtgcagc caggagggtc actgcgactg 60

agctgtgccg tgtccgggat cgacctgtca cacgacgata ttagctgggt gagacaggca 120

cccgggaagg gactggaatg ggtctctatc attagttacg ccggcagcac ctactatgcc 180

tcttgggcta aaggcaggct gaccattttct aaggacacaa gtaaaaacca ggtggtcctg 240

accatgacaa atatggaccc cgtggatact gcaacctact attgcgcccg gggcgtccct 300

gcttatagcc acggcggcga tctgtgggga caggggacac tggtgactgt cagctcc 357

<210> 107

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 107

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcaaag tgagtgggat cgacctgagc cacgacgaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtctccatc attagctacg ccgggagcac atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 108

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 108

caggtgacac tgaaagaaag cggccccgtg ctggtgaagc caaccgagac actgactctg 60

acctgcaaag tgagtgggat cgacctgagc cacgaggaca tctcctgggt gcggcaggca 120

ccgggaaagg gcctggaatg ggtctccatc attagctacg ccgggagcac atactatgcc 180

agctgggcta aaggaagact gactatctcc aaggacacct ctaaaaacca ggtggtcctg 240

actatgacca atatggaccc cgtggataca gcaacttact attgtgccag gggcgtccct 300

gcttactctc acggagggga tctgtgggga cagggaaccc tggtgacagt ctctagt 357

<210> 109

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 109

caggtgcagc tggtcgaatc tggaggagga ctggtgcagc caggagggtc actgcgactg 60

agctgtgccg tgtccgggat cgacctgtca cacgaggata ttagctgggt gagacaggca 120

cccgggaagg gactggaatg ggtctctatc attagttacg ccggcagcaa gtactatgcc 180

tcttgggcta aaggcaggct gaccattttct aaggacacaa gtaaaaacca ggtggtcctg 240

accatgacaa atatggaccc cgtggatact gcaacctact attgcgcccg gggcgtccct 300

gcttatagcc acggcggcga tctgtgggga caggggacac tggtgactgt cagctcc 357

<210> 110

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 110

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgca ccagcagcca gtccgtgtac cacgccaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc ctacggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gcggtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 111

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 111

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgca ccaccagcca gtccgtgtac cacgagaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc ctacggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gcggtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 112

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 112

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgca ccagcagcca gtccgtgttc cacgagaatt ggctgagctg gttccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc ctggggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgccgctac ctactattgt gctggcgggt acggaggcgg gcggtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 113

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 113

gacattgtga tgactcagtc tcctgccact ctgtcactga gccctggcga gcgagcaacc 60

ctgtcttgca ccaccagcca gtccgtgtac cacgagaatt ggctgagctg gtaccagcag 120

aagccagggc agccccctaa actgctgatc tattgggcat caacactggc ctacggcgtc 180

ccctccagat tctccggatc tggcagtggg actgacttta ccctgaccat cagcagcctc 240

cagcccgagg atgtcgctac ctactattgt gctggcgggt acggaggcgg gcggtatgca 300

tttggacagg gcacaaaggt ggaaatcaaa 330

<210> 114

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 114

His Asp Asp Ile Ser

15

<210> 115

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 115

His Glu Asp Ile Ser

15

<210> 116

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 116

Ile Ile Ser Tyr Ala Gly Ser Met Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 117

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 117

Ile Ile Ser Tyr Ala Gly Lys Thr Tyr Tyr Ala Ser Trp Ala Lys Lys

1 5 10 15

<210> 118

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 118

Ile Ile Ser Tyr Ala Gly Lys Lys Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 119

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 119

Ile Ile Ser Tyr Ala Gly Ser Lys Tyr Lys Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 120

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 120

Ile Ile Ser Tyr Ala Gly Ser Lys Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 121

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 121

Gly Val Pro Ala Tyr Ser His Gly Gly Asp Lys

1 5 10

<210> 122

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 122

Thr Ser Ser Gln Ser Val Tyr His Ala Asn Trp Leu Ser

1 5 10

<210> 123

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 123

Thr Thr Ser Gln Ser Val Tyr His Glu Asn Trp Leu Ser

1 5 10

<210> 124

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 124

Thr Ser Ser Gln Ser Val Phe His Glu Asn Trp Leu Ser

1 5 10

<210> 125

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 125

Trp Ala Ser Thr Leu Ala Trp

15

<210> 126

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<220>

<221> misc_feature

<222> (1)..(1)

<223> Xaa is Ser or His

<220>

<221> misc_feature

<222> (2)..(2)

<223> Xaa is Tyr, Thr, Asp or Glu

<400> 126

Xaa Xaa Asp Ile Ser

15

<210> 127

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<220>

<221> misc_feature

<222> (4)..(4)

<223> Xaa is Tyr or His

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa is Ser or Lys

<220>

<221> misc_feature

<222>(8)..(8)

<223> Xaa is Thr, Met or Lys

<220>

<221> misc_feature

<222> (10)..(10)

<223> Xaa is Tyr or Lys

<220>

<221> misc_feature

<222> (11)..(11)

<223> Xaa is Ala, Met or Glu

<220>

<221> misc_feature

<222> (12)..(12)

<223> Xaa is Ser or Glu

<220>

<221> misc_feature

<222> (16)..(16)

<223> Xaa is Gly or Lys

<400> 127

Ile Ile Ser Xaa Ala Gly Xaa Xaa Tyr Xaa Xaa Xaa Trp Ala Lys Xaa

1 5 10 15

<210> 128

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<220>

<221> misc_feature

<222> (5)..(5)

<223> Xaa is Tyr or His

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa is Thr or His

<220>

<221> misc_feature

<222> (11)..(11)

<223> Xaa is Leu or Lys

<400> 128

Gly Val Pro Ala Xaa Ser Xaa Gly Gly Asp Xaa

1 5 10

<210> 129

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<220>

<221> misc_feature

<222> (1)..(1)

<223> Xaa is Gln or Thr

<220>

<221> misc_feature

<222> (2)..(2)

<223> Xaa is Ser or Thr

<220>

<221> misc_feature

<222> (5)..(5)

<223> Xaa is Ser or Glu

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa is Tyr or Phe

<220>

<221> misc_feature

<222>(8)..(8)

<223> Xaa is Asp or His

<220>

<221> misc_feature

<222>(9)..(9)

<223> Xaa is Asn, Asp, Ala or Glu

<400> 129

Xaa Xaa Ser Gln Xaa Val Xaa Xaa Xaa Asn Trp Leu Ser

1 5 10

<210> 130

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<220>

<221> misc_feature

<222> (3)..(3)

<223> Xaa is Ser or Glu

<220>

<221> misc_feature

<222>(7)..(7)

<223> Xaa is Ser, Tyr, Phe or Trp

<400> 130

Trp Ala Xaa Thr Leu Ala Xaa

15

<210> 131

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<220>

<221> misc_feature

<222>(8)..(8)

<223> Xaa is Leu or Arg

<400> 131

Ala Gly Gly Tyr Gly Gly Gly Xaa Tyr Ala

1 5 10

<210> 132

<211> 30

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 132

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser

20 25 30

<210> 133

<211> 30

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 133

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Ile Asp Leu Ser

20 25 30

<210> 134

<211> 30

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 134

Gln Val Thr Leu Lys Glu Ser Gly Pro Val Leu Val Lys Pro Thr Glu

1 5 10 15

Thr Leu Thr Leu Thr Cys Lys Val Ser Gly Ile Asp Leu Ser

20 25 30

<210> 135

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 135

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Gly

1 5 10

<210> 136

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 136

Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser

1 5 10

<210> 137

<211> 32

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 137

Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val Val Leu Thr

1 5 10 15

Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr Cys Ala Arg

20 25 30

<210> 138

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 138

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

1 5 10

<210> 139

<211> 23

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 139

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys

20

<210> 140

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 140

Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 141

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 141

Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr

1 5 10 15

<210> 142

<211> 32

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 142

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Ala Ala Thr Tyr Tyr Cys

20 25 30

<210> 143

<211> 32

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 143

Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr

1 5 10 15

Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Val Ala Thr Tyr Tyr Cys

20 25 30

<210> 144

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 144

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

1 5 10

<210> 145

<211> 121

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 145

Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Arg Tyr Tyr

20 25 30

Val Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

35 40 45

Ile Ile Tyr Thr Gly Gly Asn Thr Asn Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Ala Ser Thr Thr Val Asp Leu Lys Met Thr

65 70 75 80

Gly Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Gly Phe Leu

85 90 95

Asp Tyr Ser His Tyr Thr Thr Gly Gly Tyr Tyr Leu Asp Leu Trp Gly

100 105 110

Pro Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 146

<211> 119

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 146

Gln Ser Leu Glu Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly Thr

1 5 10 15

Leu Thr Leu Thr Cys Lys Ala Ser Gly Ile Asp Phe Ser Ser Tyr Tyr

20 25 30

Tyr Met Cys Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Ala Cys Ile Tyr Ala Gly Ile Ser Gly Ser Thr Tyr Tyr Ala Ser Trp

50 55 60

Ala Lys Gly Arg Phe Thr Ile Ser Lys Thr Ser Ser Thr Thr Val Thr

65 70 75 80

Leu Gln Met Thr Ser Leu Thr Ala Ala Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Asp Pro Tyr His Phe Asp Tyr Gly Ser Leu Trp Gly Pro Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 147

<211> 116

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 147

Gln Ser Leu Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Arg Tyr Trp

20 25 30

Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

35 40 45

Ile Ile Ser Ser Gly Gly Thr Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr

65 70 75 80

Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp Ser

85 90 95

Arg Tyr Gly Ala Ala Arg Leu Asp Leu Trp Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 148

<211> 109

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 148

Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Tyr Ile

20 25 30

Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

35 40 45

Phe Ile Tyr Val Ser Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Arg Thr Ser Thr Thr Val Asp Leu Arg Ile Thr

65 70 75 80

Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Val Arg Thr Trp

85 90 95

Asp Leu Trp Gly Pro Gly Thr Leu Val Thr Val Ser Ser

100 105

<210> 149

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 149

Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser Ser Tyr Ala

20 25 30

Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile Gly

35 40 45

Ile Ile Tyr Gly Val Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Ala

65 70 75 80

Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Asp Asp

85 90 95

Tyr Gly Trp Gly Tyr Thr Asn Leu Trp Gly Pro Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 150

<211> 117

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 150

Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro Gly Thr Pro

1 5 10 15

Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser Ser Ser Ser

20 25 30

Met Ile Trp Val Arg Gln Ala Pro Gly Glu Gly Leu Lys Tyr Ile Gly

35 40 45

Phe Ile Ser Lys Ser Gly Tyr Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

50 55 60

Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu Lys Ile Thr

65 70 75 80

Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala Arg Gly Gly

85 90 95

Tyr Ser Gly Asp Trp Asp Tyr Phe Thr Leu Trp Gly Pro Gly Thr Leu

100 105 110

Val Thr Val Ser Ser

115

<210> 151

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 151

Gln Val Leu Thr Gln Thr Pro Ser Pro Val Ser Ala Ala Val Gly Gly

1 5 10 15

Thr Val Thr Ile Asn Cys Gln Ala Ser Gln Ser Val Tyr Asn Asn Lys

20 25 30

Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

35 40 45

Ile Tyr Tyr Ala Ser Thr Leu Pro Ser Gly Val Ser Ser Arg Phe Lys

50 55 60

Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly Val Gln

65 70 75 80

Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Glu Phe Ser Cys Gly

85 90 95

Ser Ala Asp Cys Thr Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 152

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 152

Ala Leu Val Met Thr Gln Thr Pro Ala Ser Val Ser Ala Ala Val Gly

1 5 10 15

Gly Thr Val Thr Ile Asn Cys Gln Ala Ser Gln Ser Val Tyr Gly Gly

20 25 30

Asn Glu Leu Ser Trp Tyr Gln Gln Glu Pro Gly Gln Pro Pro Lys Leu

35 40 45

Leu Ile Tyr Ser Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe

50 55 60

Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly Val

65 70 75 80

Gln Cys Asp Asp Ala Ala Ala Tyr Tyr Cys Ala Gly Tyr Arg Thr Tyr

85 90 95

Ser Asp Asp Asp Asn Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 153

<211> 112

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 153

Gln Val Leu Thr Gln Thr Pro Ala Ser Val Ser Ala Ala Val Gly Gly

1 5 10 15

Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Ser Val Ala Gly Asn Asp

20 25 30

Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

35 40 45

Ile Tyr Tyr Ala Ser Asn Leu Ala Ser Gly Val Ser Ser Arg Phe Lys

50 55 60

Gly Ser Gly Ser Gly Thr Arg Phe Thr Leu Thr Ile Ser Asp Val Gln

65 70 75 80

Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Leu Gly Asn Tyr Asp Cys Ser

85 90 95

Ser Ala Asp Cys Asp Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 154

<211> 109

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 154

Gln Val Leu Thr Gln Thr Pro Ser Ser Thr Ser Ala Ala Val Gly Gly

1 5 10 15

Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Ser Val Tyr Asp Asn Asn

20 25 30

Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Arg Leu

35 40 45

Ile Tyr Gly Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Lys

50 55 60

Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Asp Leu Glu

65 70 75 80

Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Gly Tyr Ser Gly Asn

85 90 95

Ile Tyr Pro Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 155

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 155

Gln Val Leu Thr Gln Thr Pro Ser Ser Val Ser Ala Ala Val Gly Gly

1 5 10 15

Thr Val Thr Ile Asn Cys Gln Ser Ser Gln Ser Val Tyr Asn Asn Asn

20 25 30

Tyr Leu Ala Trp Phe Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

35 40 45

Ile Tyr Asp Ala Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Lys

50 55 60

Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr Ile Ser Gly Val Gln

65 70 75 80

Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Ser Tyr Tyr Ser Ser

85 90 95

Gly Trp Tyr Arg Ala Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 156

<211> 114

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 156

Ala Asp Ile Val Met Thr Gln Thr Pro Ala Ser Val Glu Ala Ala Val

1 5 10 15

Gly Gly Thr Val Thr Ile Lys Cys Gln Ala Ser Glu Ser Ile Gly Asn

20 25 30

Ala Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Lys Leu Leu

35 40 45

Ile Tyr Gly Ala Phe Asn Leu Glu Ser Gly Val Pro Ser Arg Phe Arg

50 55 60

Gly Ser Gly Ser Gly Thr Glu Tyr Thr Leu Thr Ile Ser Asp Leu Glu

65 70 75 80

Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gly Tyr Phe Trp Ser Ser

85 90 95

Ser Thr Ile Tyr Ala Phe Trp Ala Phe Gly Gly Gly Thr Lys Val Glu

100 105 110

Ile Lys

<210> 157

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 157

Arg Tyr Tyr Val Ser

15

<210> 158

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 158

Ser Tyr Tyr Tyr Met Cys

15

<210> 159

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 159

Arg Tyr Trp Met Ser

15

<210> 160

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 160

Ser Tyr Ile Met Gly

15

<210> 161

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 161

Ser Tyr Ala Met Ser

15

<210> 162

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 162

Ser Ser Ser Met Ile

15

<210> 163

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 163

Ile Ile Tyr Thr Gly Gly Asn Thr Asn Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 164

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 164

Cys Ile Tyr Ala Gly Ile Ser Gly Ser Thr Tyr Tyr Ala Ser Trp Ala

1 5 10 15

Lys Gly

<210> 165

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 165

Ile Ile Ser Ser Gly Gly Thr Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 166

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 166

Phe Ile Tyr Val Ser Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 167

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 167

Ile Ile Tyr Gly Val Gly Ser Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 168

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 168

Phe Ile Ser Lys Ser Gly Tyr Thr Tyr Tyr Ala Ser Trp Ala Lys Gly

1 5 10 15

<210> 169

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 169

Phe Leu Asp Tyr Ser His Tyr Thr Thr Gly Gly Tyr Tyr Leu Asp Leu

1 5 10 15

<210> 170

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 170

Asp Pro Tyr His Phe Asp Tyr Gly Ser Leu

1 5 10

<210> 171

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 171

Asp Ser Arg Tyr Gly Ala Ala Arg Leu Asp Leu

1 5 10

<210> 172

<211> 4

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 172

Thr Trp Asp Leu

1

<210> 173

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 173

Asp Asp Tyr Gly Trp Gly Tyr Thr Asn Leu

1 5 10

<210> 174

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 174

Gly Gly Tyr Ser Gly Asp Trp Asp Tyr Phe Thr Leu

1 5 10

<210> 175

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 175

Gln Ala Ser Gln Ser Val Tyr Asn Asn Lys Asn Leu Ala

1 5 10

<210> 176

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 176

Gln Ala Ser Gln Ser Val Tyr Gly Gly Asn Glu Leu Ser

1 5 10

<210> 177

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 177

Gln Ser Ser Gln Ser Val Ala Gly Asn Asp Asn Leu Ala

1 5 10

<210> 178

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 178

Gln Ser Ser Gln Ser Val Tyr Asp Asn Asn Trp Leu Ala

1 5 10

<210> 179

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 179

Gln Ser Ser Gln Ser Val Tyr Asn Asn Asn Tyr Leu Ala

1 5 10

<210> 180

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 180

Gln Ala Ser Glu Ser Ile Gly Asn Ala Leu Ala

1 5 10

<210> 181

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 181

Tyr Ala Ser Thr Leu Pro Ser

15

<210> 182

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 182

Ser Ala Ser Lys Leu Ala Ser

15

<210> 183

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 183

Tyr Ala Ser Asn Leu Ala Ser

15

<210> 184

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 184

Gly Ala Ser Thr Leu Asp Ser

15

<210> 185

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 185

Asp Ala Ser Lys Leu Ala Ser

15

<210> 186

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 186

Gly Ala Phe Asn Leu Glu Ser

15

<210> 187

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 187

Gln Gly Glu Phe Ser Cys Gly Ser Ala Asp Cys Thr Ala

1 5 10

<210> 188

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 188

Ala Gly Tyr Arg Thr Tyr Ser Asp Asp Asp Asn Ala

1 5 10

<210> 189

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 189

Leu Gly Asn Tyr Asp Cys Ser Ser Ala Asp Cys Asp Ala

1 5 10

<210> 190

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 190

Gln Gly Gly Tyr Ser Gly Asn Ile Tyr Pro

1 5 10

<210> 191

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 191

Gln Gly Ser Tyr Tyr Ser Ser Gly Trp Tyr Arg Ala

1 5 10

<210> 192

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 192

Gln Gly Tyr Phe Trp Ser Ser Ser Thr Ile Tyr Ala Phe Trp Ala

1 5 10 15

<210> 193

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 193

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Arg Arg Gly Pro Lys Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 194

<211> 324

<212> DNA

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 194

cgtacggtgg ctgcaccatc tgtcttcatc ttcccgccat ctgatgagca gttgaaatct 60

ggaactgcct ctgttgtgtg cctgctgaat aacttctatc ccagagaggc caaagtacag 120

tggaaggtgg ataacgccct ccaatcgggt aactcccagg agagtgtcac agagcaggac 180

agtaaggact gcacctacag tctcagtagt accctgacgc tgtccaaagc agactacgag 240

aaacacaaag tctacgcctg cgaagtcacc catcagggcc tgtcctcgcc cgtcacaaag 300

tccttcaaca ggggagagtg ttga 324

<210> 195

<211> 330

<212> PRT

<213> Homo sapiens

<400> 195

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 196

<211> 326

<212> PRT

<213> Homo sapiens

<400> 196

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro

100 105 110

Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

115 120 125

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

130 135 140

Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly

145 150 155 160

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn

165 170 175

Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp

180 185 190

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro

195 200 205

Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu

210 215 220

Pro Gln Val Tyr Thr Leu Pro Ser Arg Glu Glu Met Thr Lys Asn

225 230 235 240

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

245 250 255

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

260 265 270

Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

275 280 285

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

290 295 300

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

305 310 315 320

Ser Leu Ser Pro Gly Lys

325

<210> 197

<211> 377

<212> PRT

<213> Homo sapiens

<400> 197

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Leu Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro

100 105 110

Arg Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg

115 120 125

Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys

130 135 140

Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys Pro

145 150 155 160

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

165 170 175

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

180 185 190

Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Lys Trp Tyr

195 200 205

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

210 215 220

Gln Tyr Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Leu His

225 230 235 240

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

245 250 255

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln

260 265 270

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Ser Arg Glu Glu Met

275 280 285

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

290 295 300

Ser Asp Ile Ala Val Glu Trp Glu Ser Ser Gly Gln Pro Glu Asn Asn

305 310 315 320

Tyr Asn Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu

325 330 335

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Ile

340 345 350

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn Arg Phe Thr Gln

355 360 365

Lys Ser Leu Ser Leu Ser Pro Gly Lys

370 375

<210> 198

<211> 327

<212> PRT

<213> Homo sapiens

<400> 198

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro

100 105 110

Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

115 120 125

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

130 135 140

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155 160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200 205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315 320

Leu Ser Leu Ser Leu Gly Lys

325

<210> 199

<211> 1125

<212> DNA

<213> Homo sapiens

<400> 199

atgtggttct tgacaactct gctcctttgg gttccagttg atgggcaagt ggacaccaca 60

aaggcagtga tcactttgca gcctccatgg gtcagcgtgt tccaagagga aaccgtaacc 120

ttgcactgtg aggtgctcca tctgcctggg agcagctcta cacagtggtt tctcaatggc 180

acagccactc agacctcgac ccccagctac agaatcacct ctgccagtgt caatgacagt 240

ggtgaataca ggtgccagag aggtctctca gggcgaagtg accccataca gctggaaatc 300

cacagaggct ggctactact gcaggtctcc agcagagtct tcacgggaagg agaacctctg 360

gccttgaggt gtcatgcgtg gaaggataag ctggtgtaca atgtgcttta ctatcgaaat 420

ggcaaagcct ttaagttttt ccactggaat tctaacctca ccattctgaa aaccaacata 480

agtcacaatg gcacctacca ttgctcaggc atgggaaagc atcgctacac atcagcagga 540

atatctgtca ctgtgaaaga gctatttcca gctccagtgc tgaatgcatc tgtgacatcc 600

ccactcctgg aggggaatct ggtcaccctg agctgtgaaa caaagttgct cttgcagagg 660

cctggtttgc agctttactt ctccttctac atgggcagca agaccctgcg aggcaggaac 720

acatcctctg aataccaaat actaactgct agaagagaag actctgggtt atactggtgc 780

gaggctgcca cagaggatgg aaatgtcctt aagcgcagcc ctgagttgga gcttcaagtg 840

cttggcctcc agttaccaac tcctgtctgg tttcatgtcc ttttctatct ggcagtggga 900

ataatgtttt tagtgaacac tgttctctgg gtgacaatac gtaaagaact gaaaagaaag 960

aaaaagtggg atttagaaat ctctttggat tctggtcatg agaagaaggt aatttccagc 1020

cttcaagaag acagacattt agaagaagag ctgaaatgtc aggaacaaaa agaagaacag 1080

ctgcaggaag gggtgcaccg gaaggagccc cagggggcca cgtag 1125

<210> 200

<211> 951

<212> DNA

<213> Homo sapiens

<400> 200

atgactatgg agacccaaat gtctcagaat gtatgtccca gaaacctgtg gctgcttcaa 60

ccattgacag ttttgctgct gctggcttct gcagacagtc aagctgctcc cccaaaggct 120

gtgctgaaac ttgagcccc gtggatcaac gtgctccagg aggactctgt gactctgaca 180

tgccaggggg ctcgcagccc tgagagcgac tccattcagt ggttccacaa tgggaatctc 240

attcccaccc acacgcagcc cagctacagg ttcaaggcca acaacaatga cagcggggag 300

tacacgtgcc agactggcca gaccagcctc agcgaccctg tgcatctgac tgtgctttcc 360

gaatggctgg tgctccagac ccctcacctg gagttccagg agggagaaac catcatgctg 420

aggtgccaca gctggaagga caagcctctg gtcaaggtca cattcttcca gaatggaaaa 480

tccgaaat tctcccattt ggatcccacc ttctccatcc cacaagcaaa ccacagtcac 540

agtggtgatt accactgcac aggaaacata ggctacacgc tgttctcatc caagcctgtg 600

accatcactg tccaagtgcc cagcatgggc agctcttcac caatgggggt cattgtggct 660

gtggtcattg cgactgctgt agcagccatt gttgctgctg tagtggcctt gatctactgc 720

aggaaaaagc ggatttcagc caattccact gatcctgtga aggctgccca atttgagcca 780

cctggacgtc aaatgattgc catcagaaag agacaacttg aagaaaccaa caatgactat 840

gaaacagctg acggcggcta catgactctg aaccccaggg cacctactga cgatgataaa 900

aacatctacc tgactcttcc tcccaacgac catgtcaaca gtaataacta a 951

<210> 201

<211> 954

<212> DNA

<213> Homo sapiens

<400> 201

atgactatgg agacccaaat gtctcagaat gtatgtccca gaaacctgtg gctgcttcaa 60

ccattgacag ttttgctgct gctggcttct gcagacagtc aagctgcagc tcccccaaag 120

gctgtgctga aacttgagcc cccgtggatc aacgtgctcc aggaggactc tgtgactctg 180

acatgccagg gggctcgcag ccctgagagc gactccattc agtggttcca caatgggaat 240

ctcattccca cccacacgca gcccagctac aggttcaagg ccaacaacaa tgacagcggg 300

gagtacacgt gccagactgg ccagaccagc ctcagcgacc ctgtgcatct gactgtgctt 360

tccgaatggc tggtgctcca gacccctcac ctggagttcc aggagggaga aaccatcatg 420

ctgaggtgcc acagctggaa ggacaagcct ctggtcaagg tcacattctt ccagaatgga 480

aaatcccaga aattctccca tttggatccc accttctcca tcccacaagc aaaccacagt 540

cacagtggtg attaccactg cacaggaaac ataggctaca cgctgttctc atccaagcct 600

gtgaccatca ctgtccaagt gcccagcatg ggcagctctt caccaatggg gatcattgtg 660

gctgtggtca ttgcgactgc tgtagcagcc attgttgctg ctgtagtggc cttgatctac 720

tgcaggaaaa agcggatttc agccaattcc actgatcctg tgaaggctgc ccaatttgag 780

ccacctggac gtcaaatgat tgccatcaga aagagacaac ttgaagaaac caacaatgac 840

tatgaaacag ctgacggcgg ctacatgact ctgaacccca gggcacctac tgacgatgat 900

aaaaacatct acctgactct tcctcccaac gaccatgtca acagtaataa ctaa 954

<210> 202

<211> 876

<212> DNA

<213> Homo sapiens

<400> 202

atgggaatcc tgtcattctt acctgtcctt gccactgaga gtgactgggc tgactgcaag 60

tccccccagc cttggggtca tatgcttctg tggacagctg tgctattcct ggctcctgtt 120

gctgggacac ctgcagctcc cccaaaggct gtgctgaaac tcgagcccca gtggatcaac 180

gtgctccagg aggactctgt gactctgaca tgccggggga ctcacagccc tgagagcgac 240

tccattcagt ggttccacaa tgggaatctc attcccaccc acacgcagcc cagctacagg 300

ttcaaggcca acaacaatga cagcggggag tacacgtgcc agactggcca gaccagcctc 360

agcgaccctg tgcatctgac tgtgctttct gagtggctgg tgctccagac ccctcacctg 420

gagttccagg agggagaaac catcgtgctg aggtgccaca gctggaagga caagcctctg 480

gtcaaggtca cattcttcca gaatggaaaa tccaagaaat tttcccgttc ggatcccaac 540

ttctccatcc cacaagcaaa ccacagtcac agtggtgatt accactgcac aggaaacata 600

ggctacacgc tgtactcatc caagcctgtg accatcactg tccaagctcc cagctcttca 660

ccgatgggga tcattgtggc tgtggtcact gggattgctg tagcggccat tgttgctgct 720

gtagtggcct tgatctactg caggaaaaag cggatttcag ccaatcccac taatcctgat 780

gaggctgaca aagttggggc tgagaacaca atcacctatt cacttctcat gcacccggat 840

gctctggaag agcctgatga ccagaaccgt atttag 876

<210> 203

<211> 933

<212> DNA

<213> Homo sapiens

<400> 203

atgggaatcc tgtcattctt acctgtcctt gccactgaga gtgactgggc tgactgcaag 60

tccccccagc cttggggtca tatgcttctg tggacagctg tgctattcct ggctcctgtt 120

gctgggacac ctgcagctcc cccaaaggct gtgctgaaac tcgagcccca gtggatcaac 180

gtgctccagg aggactctgt gactctgaca tgccggggga ctcacagccc tgagagcgac 240

tccattcagt ggttccacaa tgggaatctc attcccaccc acacgcagcc cagctacagg 300

ttcaaggcca acaacaatga cagcggggag tacacgtgcc agactggcca gaccagcctc 360

agcgaccctg tgcatctgac tgtgctttct gagtggctgg tgctccagac ccctcacctg 420

gagttccagg agggagaaac catcgtgctg aggtgccaca gctggaagga caagcctctg 480

gtcaaggtca cattcttcca gaatggaaaa tccaagaaat tttcccgttc ggatcccaac 540

ttctccatcc cacaagcaaa ccacagtcac agtggtgatt accactgcac aggaaacata 600

ggctacacgc tgtactcatc caagcctgtg accatcactg tccaagctcc cagctcttca 660

ccgatgggga tcattgtggc tgtggtcact gggattgctg tagcggccat tgttgctgct 720

gtagtggcct tgatctactg caggaaaaag cggatttcag ctctcccagg ataccctgag 780

tgcagggaaa tgggagagac cctccctgag aaaccagcca atcccactaa tcctgatgag 840

gctgacaaag ttggggctga gaacacaatc acctattcac ttctcatgca cccggatgct 900

ctggaagagc ctgatgacca gaaccgtatt tag 933

<210> 204

<211> 765

<212> DNA

<213> Homo sapiens

<400> 204

atgtggcagc tgctcctccc aactgctctg ctacttctag tttcagctgg catgcggact 60

gaagatctcc caaaggctgt ggtgttcctg gagcctcaat ggtacagggt gctcgagaag 120

gacagtgtga ctctgaagtg ccagggagcc tactcccctg aggacaattc cacacagtgg 180

tttcacaatg agagcctcat ctcaagccag gcctcgagct acttcattga cgctgccaca 240

gttgacgaca gtggagagta caggtgccag acaaacctct ccaccctcag tgacccggtg 300

cagctagaag tccatatcgg ctggctgttg ctccaggccc ctcggtgggt gttcaaggag 360

gaagacccta ttcacctgag gtgtcacagc tggaagaaca ctgctctgca taaggtcaca 420

tatttacaga atggcaaagg caggaagtat tttcatcata attctgactt ctacattcca 480

aaagccacac tcaaagacag cggctcctac ttctgcaggg ggcttgttgg gagtaaaaat 540

gtgtcttcag agactgtgaa catcaccatc actcaaggtt tgtcagtgtc aaccatctca 600

tcattctttc cacctgggta ccaagtctct ttctgcttgg tgatggtact cctttttgca 660

gtggacacag gactatattt ctctgtgaag acaaacattc gaagctcaac aagagactgg 720

aaggaccata aatttaaatg gagaaaggac cctcaagaca aatga 765

<210> 205

<211> 765

<212> DNA

<213> Homo sapiens

<400> 205

atgtggcagc tgctcctccc aactgctctg ctacttctag tttcagctgg catgcggact 60

gaagatctcc caaaggctgt ggtgttcctg gagcctcaat ggtacagggt gctcgagaag 120

gacagtgtga ctctgaagtg ccagggagcc tactcccctg aggacaattc cacacagtgg 180

tttcacaatg agagcctcat ctcaagccag gcctcgagct acttcattga cgctgccaca 240

gtcgacgaca gtggagagta caggtgccag acaaacctct ccaccctcag tgacccggtg 300

cagctagaag tccatatcgg ctggctgttg ctccaggccc ctcggtgggt gttcaaggag 360

gaagacccta ttcacctgag gtgtcacagc tggaagaaca ctgctctgca taaggtcaca 420

tatttacaga atggcaaagg caggaagtat tttcatcata attctgactt ctacattcca 480

aaagccacac tcaaagacag cggctcctac ttctgcaggg ggctttttgg gagtaaaaat 540

gtgtcttcag agactgtgaa catcaccatc actcaaggtt tggcagtgtc aaccatctca 600

tcattctttc cacctgggta ccaagtctct ttctgcttgg tgatggtact cctttttgca 660

gtggacacag gactatattt ctctgtgaag acaaacattc gaagctcaac aagagactgg 720

aaggaccata aatttaaatg gagaaaggac cctcaagaca aatga 765

<210> 206

<211> 702

<212> DNA

<213> Homo sapiens

<400> 206

atgtggcagc tgctcctccc aactgctctg ctacttctag tttcagctgg catgcggact 60

gaagatctcc caaaggctgt ggtgttcctg gagcctcaat ggtacagcgt gcttgagaag 120

gacagtgtga ctctgaagtg ccagggagcc tactcccctg aggacaattc cacacagtgg 180

tttcacaatg agagcctcat ctcaagccag gcctcgagct acttcattga cgctgccaca 240

gtcaacgaca gtggagagta caggtgccag acaaacctct ccaccctcag tgacccggtg 300

cagctagaag tccatatcgg ctggctgttg ctccaggccc ctcggtgggt gttcaaggag 360

gaagacccta ttcacctgag gtgtcacagc tggaagaaca ctgctctgca taaggtcaca 420

tatttacaga atggcaaaga caggaagtat tttcatcata attctgactt ccacattcca 480

aaagccacac tcaaagatag cggctcctac ttctgcaggg ggcttgttgg gagtaaaaat 540

gtgtcttcag agactgtgaa catcaccatc actcaaggtt tggcagtgtc aaccatctca 600

tcattctctc cacctgggta ccaagtctct ttctgcttgg tgatggtact cctttttgca 660

gtggacacag gactatattt ctctgtgaag acaaacattt ga 702

<210> 207

<211> 374

<212> PRT

<213> Homo sapiens

<400> 207

Met Trp Phe Leu Thr Thr Leu Leu Leu Trp Val Pro Val Asp Gly Gln

1 5 10 15

Val Asp Thr Thr Lys Ala Val Ile Thr Leu Gln Pro Pro Trp Val Ser

20 25 30

Val Phe Gln Glu Glu Thr Val Thr Leu His Cys Glu Val Leu His Leu

35 40 45

Pro Gly Ser Ser Ser Thr Gln Trp Phe Leu Asn Gly Thr Ala Thr Gln

50 55 60

Thr Ser Thr Pro Ser Tyr Arg Ile Thr Ser Ala Ser Val Asn Asp Ser

65 70 75 80

Gly Glu Tyr Arg Cys Gln Arg Gly Leu Ser Gly Arg Ser Asp Pro Ile

85 90 95

Gln Leu Glu Ile His Arg Gly Trp Leu Leu Leu Gln Val Ser Ser Arg

100 105 110

Val Phe Thr Glu Gly Glu Pro Leu Ala Leu Arg Cys His Ala Trp Lys

115 120 125

Asp Lys Leu Val Tyr Asn Val Leu Tyr Tyr Arg Asn Gly Lys Ala Phe

130 135 140

Lys Phe Phe His Trp Asn Ser Asn Leu Thr Ile Leu Lys Thr Asn Ile

145 150 155 160

Ser His Asn Gly Thr Tyr His Cys Ser Gly Met Gly Lys His Arg Tyr

165 170 175

Thr Ser Ala Gly Ile Ser Val Thr Val Lys Glu Leu Phe Pro Ala Pro

180 185 190

Val Leu Asn Ala Ser Val Thr Ser Pro Leu Leu Glu Gly Asn Leu Val

195 200 205

Thr Leu Ser Cys Glu Thr Lys Leu Leu Leu Gln Arg Pro Gly Leu Gln

210 215 220

Leu Tyr Phe Ser Phe Tyr Met Gly Ser Lys Thr Leu Arg Gly Arg Asn

225 230 235 240

Thr Ser Ser Glu Tyr Gln Ile Leu Thr Ala Arg Arg Glu Asp Ser Gly

245 250 255

Leu Tyr Trp Cys Glu Ala Ala Thr Glu Asp Gly Asn Val Leu Lys Arg

260 265 270

Ser Pro Glu Leu Glu Leu Gln Val Leu Gly Leu Gln Leu Pro Thr Pro

275 280 285

Val Trp Phe His Val Leu Phe Tyr Leu Ala Val Gly Ile Met Phe Leu

290 295 300

Val Asn Thr Val Leu Trp Val Thr Ile Arg Lys Glu Leu Lys Arg Lys

305 310 315 320

Lys Lys Trp Asp Leu Glu Ile Ser Leu Asp Ser Gly His Glu Lys Lys

325 330 335

Val Ile Ser Ser Leu Gln Glu Asp Arg His Leu Glu Glu Glu Glu Leu Lys

340 345 350

Cys Gln Glu Gln Lys Glu Glu Gln Leu Gln Glu Gly Val His Arg Lys

355 360 365

Glu Pro Gln Gly Ala Thr

370

<210> 208

<211> 316

<212> PRT

<213> Homo sapiens

<400> 208

Met Thr Met Glu Thr Gln Met Ser Gln Asn Val Cys Pro Arg Asn Leu

1 5 10 15

Trp Leu Leu Gln Pro Leu Thr Val Leu Leu Leu Leu Ala Ser Ala Asp

20 25 30

Ser Gln Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Pro Trp

35 40 45

Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Gln Gly Ala

50 55 60

Arg Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn Leu

65 70 75 80

Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn

85 90 95

Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser Asp

100 105 110

Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr Pro

115 120 125

His Leu Glu Phe Gln Glu Gly Glu Thr Ile Met Leu Arg Cys His Ser

130 135 140

Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly Lys

145 150 155 160

Ser Gln Lys Phe Ser His Leu Asp Pro Thr Phe Ser Ile Pro Gln Ala

165 170 175

Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr

180 185 190

Thr Leu Phe Ser Ser Lys Pro Val Thr Ile Thr Val Gln Val Pro Ser

195 200 205

Met Gly Ser Ser Ser Pro Met Gly Val Ile Val Ala Val Val Ile Ala

210 215 220

Thr Ala Val Ala Ala Ile Val Ala Ala Val Val Ala Leu Ile Tyr Cys

225 230 235 240

Arg Lys Lys Arg Ile Ser Ala Asn Ser Thr Asp Pro Val Lys Ala Ala

245 250 255

Gln Phe Glu Pro Pro Gly Arg Gln Met Ile Ala Ile Arg Lys Arg Gln

260 265 270

Leu Glu Glu Thr Asn Asn Asp Tyr Glu Thr Ala Asp Gly Gly Tyr Met

275 280 285

Thr Leu Asn Pro Arg Ala Pro Thr Asp Asp Asp Lys Asn Ile Tyr Leu

290 295 300

Thr Leu Pro Pro Asn Asp His Val Asn Ser Asn Asn

305 310 315

<210> 209

<211> 317

<212> PRT

<213> Homo sapiens

<400> 209

Met Thr Met Glu Thr Gln Met Ser Gln Asn Val Cys Pro Arg Asn Leu

1 5 10 15

Trp Leu Leu Gln Pro Leu Thr Val Leu Leu Leu Leu Ala Ser Ala Asp

20 25 30

Ser Gln Ala Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Pro

35 40 45

Trp Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Gln Gly

50 55 60

Ala Arg Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn

65 70 75 80

Leu Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn

85 90 95

Asn Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser

100 105 110

Asp Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr

115 120 125

Pro His Leu Glu Phe Gln Glu Gly Glu Thr Ile Met Leu Arg Cys His

130 135 140

Ser Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly

145 150 155 160

Lys Ser Gln Lys Phe Ser His Leu Asp Pro Thr Phe Ser Ile Pro Gln

165 170 175

Ala Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly

180 185 190

Tyr Thr Leu Phe Ser Ser Lys Pro Val Thr Ile Thr Val Gln Val Pro

195 200 205

Ser Met Gly Ser Ser Ser Pro Met Gly Ile Ile Val Ala Val Val Ile

210 215 220

Ala Thr Ala Val Ala Ala Ile Val Ala Ala Val Val Ala Leu Ile Tyr

225 230 235 240

Cys Arg Lys Lys Arg Ile Ser Ala Asn Ser Thr Asp Pro Val Lys Ala

245 250 255

Ala Gln Phe Glu Pro Pro Gly Arg Gln Met Ile Ala Ile Arg Lys Arg

260 265 270

Gln Leu Glu Glu Thr Asn Asn Asp Tyr Glu Thr Ala Asp Gly Gly Tyr

275 280 285

Met Thr Leu Asn Pro Arg Ala Pro Thr Asp Asp Asp Lys Asn Ile Tyr

290 295 300

Leu Thr Leu Pro Pro Asn Asp His Val Asn Ser Asn Asn

305 310 315

<210> 210

<211> 218

<212> PRT

<213> Homo sapiens

<400> 210

Met Thr Met Glu Thr Gln Met Ser Gln Asn Val Cys Pro Arg Asn Leu

1 5 10 15

Trp Leu Leu Gln Pro Leu Thr Val Leu Leu Leu Leu Ala Ser Ala Asp

20 25 30

Ser Gln Ala Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Pro

35 40 45

Trp Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Gln Gly

50 55 60

Ala Arg Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn

65 70 75 80

Leu Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn

85 90 95

Asn Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser

100 105 110

Asp Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr

115 120 125

Pro His Leu Glu Phe Gln Glu Gly Glu Thr Ile Met Leu Arg Cys His

130 135 140

Ser Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly

145 150 155 160

Lys Ser Gln Lys Phe Ser Arg Leu Asp Pro Thr Phe Ser Ile Pro Gln

165 170 175

Ala Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly

180 185 190

Tyr Thr Leu Phe Ser Ser Lys Pro Val Thr Ile Thr Val Gln Val Pro

195 200 205

Ser Met Gly Ser Ser Ser Pro Met Gly Ile

210 215

<210> 211

<211> 218

<212> PRT

<213> Homo sapiens

<400> 211

Met Thr Met Glu Thr Gln Met Ser Gln Asn Val Cys Pro Arg Asn Leu

1 5 10 15

Trp Leu Leu Gln Pro Leu Thr Val Leu Leu Leu Leu Ala Ser Ala Asp

20 25 30

Ser Gln Ala Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Pro

35 40 45

Trp Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Gln Gly

50 55 60

Ala Arg Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn

65 70 75 80

Leu Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn

85 90 95

Asn Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser

100 105 110

Asp Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr

115 120 125

Pro His Leu Glu Phe Gln Glu Gly Glu Thr Ile Met Leu Arg Cys His

130 135 140

Ser Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly

145 150 155 160

Lys Ser Gln Lys Phe Ser His Leu Asp Pro Thr Phe Ser Ile Pro Gln

165 170 175

Ala Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly

180 185 190

Tyr Thr Leu Phe Ser Ser Lys Pro Val Thr Ile Thr Val Gln Val Pro

195 200 205

Ser Met Gly Ser Ser Ser Pro Met Gly Ile

210 215

<210> 212

<211> 291

<212> PRT

<213> Homo sapiens

<400> 212

Met Gly Ile Leu Ser Phe Leu Pro Val Leu Ala Thr Glu Ser Asp Trp

1 5 10 15

Ala Asp Cys Lys Ser Pro Gln Pro Trp Gly His Met Leu Leu Trp Thr

20 25 30

Ala Val Leu Phe Leu Ala Pro Val Ala Gly Thr Pro Ala Ala Pro Pro

35 40 45

Lys Ala Val Leu Lys Leu Glu Pro Gln Trp Ile Asn Val Leu Gln Glu

50 55 60

Asp Ser Val Thr Leu Thr Cys Arg Gly Thr His Ser Pro Glu Ser Asp

65 70 75 80

Ser Ile Gln Trp Phe His Asn Gly Asn Leu Ile Pro Thr His Thr Gln

85 90 95

Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn Asp Ser Gly Glu Tyr Thr

100 105 110

Cys Gln Thr Gly Gln Thr Ser Leu Ser Asp Pro Val His Leu Thr Val

115 120 125

Leu Ser Glu Trp Leu Val Leu Gln Thr Pro His Leu Glu Phe Gln Glu

130 135 140

Gly Glu Thr Ile Val Leu Arg Cys His Ser Trp Lys Asp Lys Pro Leu

145 150 155 160

Val Lys Val Thr Phe Phe Gln Asn Gly Lys Ser Lys Lys Phe Ser Arg

165 170 175

Ser Asp Pro Asn Phe Ser Ile Pro Gln Ala Asn His Ser His Ser Gly

180 185 190

Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr Thr Leu Tyr Ser Ser Lys

195 200 205

Pro Val Thr Ile Thr Val Gln Ala Pro Ser Ser Ser Pro Met Gly Ile

210 215 220

Ile Val Ala Val Val Thr Gly Ile Ala Val Ala Ala Ile Val Ala Ala

225 230 235 240

Val Val Ala Leu Ile Tyr Cys Arg Lys Lys Arg Ile Ser Ala Asn Pro

245 250 255

Thr Asn Pro Asp Glu Ala Asp Lys Val Gly Ala Glu Asn Thr Ile Thr

260 265 270

Tyr Ser Leu Leu Met His Pro Asp Ala Leu Glu Glu Pro Asp Asp Gln

275 280 285

Asn Arg Ile

290

<210> 213

<211> 310

<212> PRT

<213> Homo sapiens

<400> 213

Met Gly Ile Leu Ser Phe Leu Pro Val Leu Ala Thr Glu Ser Asp Trp

1 5 10 15

Ala Asp Cys Lys Ser Pro Gln Pro Trp Gly His Met Leu Leu Trp Thr

20 25 30

Ala Val Leu Phe Leu Ala Pro Val Ala Gly Thr Pro Ala Ala Pro Pro

35 40 45

Lys Ala Val Leu Lys Leu Glu Pro Gln Trp Ile Asn Val Leu Gln Glu

50 55 60

Asp Ser Val Thr Leu Thr Cys Arg Gly Thr His Ser Pro Glu Ser Asp

65 70 75 80

Ser Ile Gln Trp Phe His Asn Gly Asn Leu Ile Pro Thr His Thr Gln

85 90 95

Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn Asp Ser Gly Glu Tyr Thr

100 105 110

Cys Gln Thr Gly Gln Thr Ser Leu Ser Asp Pro Val His Leu Thr Val

115 120 125

Leu Ser Glu Trp Leu Val Leu Gln Thr Pro His Leu Glu Phe Gln Glu

130 135 140

Gly Glu Thr Ile Val Leu Arg Cys His Ser Trp Lys Asp Lys Pro Leu

145 150 155 160

Val Lys Val Thr Phe Phe Gln Asn Gly Lys Ser Lys Lys Phe Ser Arg

165 170 175

Ser Asp Pro Asn Phe Ser Ile Pro Gln Ala Asn His Ser His Ser Gly

180 185 190

Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr Thr Leu Tyr Ser Ser Lys

195 200 205

Pro Val Thr Ile Thr Val Gln Ala Pro Ser Ser Ser Pro Met Gly Ile

210 215 220

Ile Val Ala Val Val Thr Gly Ile Ala Val Ala Ala Ile Val Ala Ala

225 230 235 240

Val Val Ala Leu Ile Tyr Cys Arg Lys Lys Arg Ile Ser Ala Leu Pro

245 250 255

Gly Tyr Pro Glu Cys Arg Glu Met Gly Glu Thr Leu Pro Glu Lys Pro

260 265 270

Ala Asn Pro Thr Asn Pro Asp Glu Ala Asp Lys Val Gly Ala Glu Asn

275 280 285

Thr Ile Thr Tyr Ser Leu Leu Met His Pro Asp Ala Leu Glu Glu Pro

290 295 300

Asp Asp Gln Asn Arg Ile

305 310

<210> 214

<211> 217

<212> PRT

<213> Homo sapiens

<400> 214

Met Gly Ile Leu Ser Phe Leu Pro Val Leu Ala Thr Glu Ser Asp Trp

1 5 10 15

Ala Asp Cys Lys Ser Pro Gln Pro Trp Gly His Met Leu Leu Trp Thr

20 25 30

Ala Val Leu Phe Leu Ala Pro Val Ala Gly Thr Pro Ala Ala Pro Pro

35 40 45

Lys Ala Val Leu Lys Leu Glu Pro Gln Trp Ile Asn Val Leu Gln Glu

50 55 60

Asp Ser Val Thr Leu Thr Cys Arg Gly Thr His Ser Pro Glu Ser Asp

65 70 75 80

Ser Ile Gln Trp Phe His Asn Gly Asn Leu Ile Pro Thr His Thr Gln

85 90 95

Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn Asp Ser Gly Glu Tyr Thr

100 105 110

Cys Gln Thr Gly Gln Thr Ser Leu Ser Asp Pro Val His Leu Thr Val

115 120 125

Leu Ser Glu Trp Leu Val Leu Gln Thr Pro His Leu Glu Phe Gln Glu

130 135 140

Gly Glu Thr Ile Val Leu Arg Cys His Ser Trp Lys Asp Lys Pro Leu

145 150 155 160

Val Lys Val Thr Phe Phe Gln Asn Gly Lys Ser Lys Lys Phe Ser Arg

165 170 175

Ser Asp Pro Asn Phe Ser Ile Pro Gln Ala Asn His Ser His Ser Gly

180 185 190

Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr Thr Leu Tyr Ser Ser Lys

195 200 205

Pro Val Thr Ile Thr Val Gln Ala Pro

210 215

<210> 215

<211> 254

<212> PRT

<213> Homo sapiens

<400> 215

Met Trp Gln Leu Leu Leu Pro Thr Ala Leu Leu Leu Leu Val Ser Ala

1 5 10 15

Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

20 25 30

Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

35 40 45

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu

50 55 60

Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr

65 70 75 80

Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu

85 90 95

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

100 105 110

Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys

115 120 125

His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn

130 135 140

Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro

145 150 155 160

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Val

165 170 175

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

180 185 190

Gly Leu Ser Val Ser Thr Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

195 200 205

Val Ser Phe Cys Leu Val Met Val Leu Leu Phe Ala Val Asp Thr Gly

210 215 220

Leu Tyr Phe Ser Val Lys Thr Asn Ile Arg Ser Ser Thr Arg Asp Trp

225 230 235 240

Lys Asp His Lys Phe Lys Trp Arg Lys Asp Pro Gln Asp Lys

245 250

<210> 216

<211> 254

<212> PRT

<213> Homo sapiens

<400> 216

Met Trp Gln Leu Leu Leu Pro Thr Ala Leu Leu Leu Leu Val Ser Ala

1 5 10 15

Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

20 25 30

Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

35 40 45

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu

50 55 60

Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr

65 70 75 80

Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu

85 90 95

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

100 105 110

Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys

115 120 125

His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn

130 135 140

Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro

145 150 155 160

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Phe

165 170 175

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

180 185 190

Gly Leu Ala Val Ser Thr Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

195 200 205

Val Ser Phe Cys Leu Val Met Val Leu Leu Phe Ala Val Asp Thr Gly

210 215 220

Leu Tyr Phe Ser Val Lys Thr Asn Ile Arg Ser Ser Thr Arg Asp Trp

225 230 235 240

Lys Asp His Lys Phe Lys Trp Arg Lys Asp Pro Gln Asp Lys

245 250

<210> 217

<211> 208

<212> PRT

<213> Homo sapiens

<400> 217

Met Trp Gln Leu Leu Leu Pro Thr Ala Leu Leu Leu Leu Val Ser Ala

1 5 10 15

Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

20 25 30

Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

35 40 45

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu

50 55 60

Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr

65 70 75 80

Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu

85 90 95

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

100 105 110

Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys

115 120 125

His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn

130 135 140

Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro

145 150 155 160

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Phe

165 170 175

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

180 185 190

Gly Leu Ala Val Ser Thr Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

195 200 205

<210> 218

<211> 208

<212> PRT

<213> Homo sapiens

<400> 218

Met Trp Gln Leu Leu Leu Pro Thr Ala Leu Leu Leu Leu Val Ser Ala

1 5 10 15

Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

20 25 30

Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

35 40 45

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu

50 55 60

Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr

65 70 75 80

Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu

85 90 95

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

100 105 110

Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys

115 120 125

His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn

130 135 140

Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro

145 150 155 160

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Val

165 170 175

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

180 185 190

Gly Leu Ala Val Ser Thr Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

195 200 205

<210> 219

<211> 233

<212> PRT

<213> Homo sapiens

<400> 219

Met Trp Gln Leu Leu Leu Pro Thr Ala Leu Leu Leu Leu Val Ser Ala

1 5 10 15

Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

20 25 30

Gln Trp Tyr Ser Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

35 40 45

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu

50 55 60

Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr

65 70 75 80

Val Asn Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu

85 90 95

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

100 105 110

Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys

115 120 125

His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn

130 135 140

Gly Lys Asp Arg Lys Tyr Phe His His Asn Ser Asp Phe His Ile Pro

145 150 155 160

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Val

165 170 175

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

180 185 190

Gly Leu Ala Val Ser Thr Ile Ser Ser Phe Ser Pro Pro Gly Tyr Gln

195 200 205

Val Ser Phe Cys Leu Val Met Val Leu Leu Phe Ala Val Asp Thr Gly

210 215 220

Leu Tyr Phe Ser Val Lys Thr Asn Ile

225 230

<210> 220

<211> 211

<212> PRT

<213> Macaca fascicularis

<400> 220

Met Ser Gln Asn Val Cys Pro Gly Asn Leu Trp Leu Leu Gln Pro Leu

1 5 10 15

Thr Val Leu Leu Leu Leu Ala Ser Ala Asp Ser Gln Thr Ala Pro Pro

20 25 30

Lys Ala Val Leu Lys Leu Glu Pro Pro Trp Ile Asn Val Leu Arg Glu

35 40 45

Asp Ser Val Thr Leu Thr Cys Gly Gly Ala His Ser Pro Asp Ser Asp

50 55 60

Ser Thr Gln Trp Phe His Asn Gly Asn Leu Ile Pro Thr His Thr Gln

65 70 75 80

Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn Asp Ser Gly Glu Tyr Arg

85 90 95

Cys Gln Thr Gly Arg Thr Ser Leu Ser Asp Pro Val His Leu Thr Val

100 105 110

Leu Ser Glu Trp Leu Ala Leu Gln Thr Pro His Leu Glu Phe Arg Glu

115 120 125

Gly Glu Thr Ile Met Leu Arg Cys His Ser Trp Lys Asp Lys Pro Leu

130 135 140

Ile Lys Val Thr Phe Phe Gln Asn Gly Ile Ala Lys Lys Phe Ser His

145 150 155 160

Met Asp Pro Asn Phe Ser Ile Pro Arg Ala Asn His Ser His Ser Gly

165 170 175

Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr Thr Pro Tyr Ser Ser Lys

180 185 190

Pro Val Thr Ile Thr Val Gln Val Pro Ser Val Gly Ser Ser Ser Pro

195 200 205

Met Gly Ile

210

<210> 221

<211> 211

<212> PRT

<213> Macaca fascicularis

<400> 221

Met Ser Gln Asn Val Cys Pro Gly Asn Leu Trp Leu Leu Gln Pro Leu

1 5 10 15

Thr Val Leu Leu Leu Leu Ala Ser Ala Asp Ser Gln Thr Ala Pro Pro

20 25 30

Lys Ala Val Leu Lys Leu Glu Pro Pro Trp Ile Asn Val Leu Arg Glu

35 40 45

Asp Ser Val Thr Leu Thr Cys Gly Gly Ala His Ser Pro Asp Ser Asp

50 55 60

Ser Thr Gln Trp Phe His Asn Gly Asn Leu Ile Pro Thr His Thr Gln

65 70 75 80

Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn Asp Ser Gly Glu Tyr Arg

85 90 95

Cys Gln Thr Gly Arg Thr Ser Leu Ser Asp Pro Val His Leu Thr Val

100 105 110

Leu Ser Glu Trp Leu Ala Leu Gln Thr Thr His Leu Glu Phe Arg Glu

115 120 125

Gly Glu Thr Ile Met Leu Arg Cys His Ser Trp Lys Asp Lys Pro Leu

130 135 140

Ile Lys Val Ala Phe Phe Gln Asn Gly Lys Ser Lys Asn Phe Ser His

145 150 155 160

Met Asn Pro Asn Phe Ser Ile Pro Gln Ala Asn His Ser His Ser Gly

165 170 175

Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr Thr Pro Tyr Ser Ser Lys

180 185 190

Pro Val Thr Ile Thr Val Gln Val Pro Ser Val Gly Ser Ser Ser Pro

195 200 205

Met Gly Ile

210

<210> 222

<211> 211

<212> PRT

<213> Macaca fascicularis

<400> 222

Met Ser Gln Asn Val Cys Pro Gly Asn Leu Trp Leu Leu Gln Pro Leu

1 5 10 15

Thr Val Leu Leu Leu Leu Ala Ser Ala Asp Ser Gln Thr Ala Pro Pro

20 25 30

Lys Ala Val Leu Lys Leu Glu Pro Pro Trp Ile Asn Val Leu Arg Glu

35 40 45

Asp Ser Val Thr Leu Thr Cys Gly Gly Ala His Ser Pro Asp Ser Asp

50 55 60

Ser Thr Gln Trp Phe His Asn Gly Asn Leu Ile Pro Thr His Thr Gln

65 70 75 80

Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn Asp Ser Gly Glu Tyr Arg

85 90 95

Cys Gln Thr Gly Arg Thr Ser Leu Ser Asp Pro Val His Leu Thr Val

100 105 110

Leu Ser Glu Trp Leu Ala Leu Gln Thr Thr His Leu Glu Phe Arg Glu

115 120 125

Gly Glu Thr Ile Met Leu Arg Cys His Ser Trp Lys Asp Lys Pro Leu

130 135 140

Ile Lys Val Ala Phe Phe Gln Asn Gly Ile Ser Lys Lys Phe Ser Pro

145 150 155 160

Met Asn Pro Asn Phe Ser Ile Pro Arg Ala Asn His Ser His Ser Gly

165 170 175

Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr Thr Pro Tyr Ser Ser Lys

180 185 190

Pro Val Thr Ile Thr Val Gln Val Pro Ser Val Gly Ser Ser Ser Pro

195 200 205

Met Gly Ile

210

<210> 223

<211> 217

<212> PRT

<213> Macaca fascicularis

<400> 223

Met Gly Ile Leu Ser Phe Leu Pro Val Leu Ala Thr Glu Ser Asp Trp

1 5 10 15

Ala Asp Cys Lys Ser Ser Gln Pro Trp Gly His Met Leu Leu Trp Thr

20 25 30

Ala Val Leu Phe Leu Ala Pro Val Ala Gly Thr Pro Ala Ala Pro Pro

35 40 45

Lys Ala Val Leu Lys Leu Glu Pro Pro Trp Ile Asn Val Leu Arg Glu

50 55 60

Asp Ser Val Thr Leu Thr Cys Gly Gly Ala His Ser Pro Asp Ser Asp

65 70 75 80

Ser Thr Gln Trp Phe His Asn Gly Asn Leu Ile Pro Thr His Thr Gln

85 90 95

Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn Asp Ser Gly Glu Tyr Arg

100 105 110

Cys Gln Thr Gly Arg Thr Ser Leu Ser Asp Pro Val His Leu Thr Val

115 120 125

Leu Ser Glu Trp Leu Ala Leu Gln Thr Pro His Leu Glu Phe Arg Glu

130 135 140

Gly Glu Thr Ile Met Leu Arg Cys His Ser Trp Lys Asp Lys Pro Leu

145 150 155 160

Ile Lys Val Thr Phe Phe Gln Asn Gly Ile Ser Lys Lys Phe Ser His

165 170 175

Met Asn Pro Asn Phe Ser Ile Pro Gln Ala Asn His Ser His Ser Gly

180 185 190

Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr Thr Pro Tyr Ser Ser Lys

195 200 205

Pro Val Thr Ile Thr Val Gln Val Pro

210 215

<210> 224

<211> 208

<212> PRT

<213> Macaca fascicularis

<400> 224

Met Trp Gln Leu Leu Leu Pro Thr Ala Leu Leu Leu Leu Val Ser Ala

1 5 10 15

Gly Met Arg Ala Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

20 25 30

Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

35 40 45

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Arg Trp Phe His Asn Glu

50 55 60

Ser Leu Ile Ser Ser Gln Thr Ser Ser Tyr Phe Ile Ala Ala Ala Arg

65 70 75 80

Val Asn Asn Ser Gly Glu Tyr Arg Cys Gln Thr Ser Leu Ser Thr Leu

85 90 95

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

100 105 110

Ala Pro Arg Trp Val Phe Lys Glu Glu Glu Ser Ile His Leu Arg Cys

115 120 125

His Ser Trp Lys Asn Thr Leu Leu His Lys Val Thr Tyr Leu Gln Asn

130 135 140

Gly Lys Gly Arg Lys Tyr Phe His Gln Asn Ser Asp Phe Tyr Ile Pro

145 150 155 160

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Ile

165 170 175

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

180 185 190

Asp Leu Ala Val Ser Ser Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

195 200 205

<210> 225

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 225

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Thr Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Glu Met His Trp Ile Arg Gln Pro Pro Gly Glu Gly Leu Glu Trp Ile

35 40 45

Gly Ala Ile Asp Pro Lys Thr Gly Asp Thr Ala Tyr Ser Glu Ser Phe

50 55 60

Gln Asp Arg Val Thr Leu Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Arg Phe Tyr Ser Tyr Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 226

<211> 219

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 226

Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Gln Ala Ser Glu Ser Leu Val His Ser

20 25 30

Asn Arg Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Asn

85 90 95

Thr His Val Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Glu

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 227

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 227

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 228

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 228

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Glu Ser Leu Ser Leu Ser Pro

325

<210> 229

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 229

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Glu Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Met Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 230

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 230

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Glu Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Met Val Leu Asp Arg Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 231

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 231

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 232

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 232

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 233

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 233

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 234

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 234

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 235

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 235

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Thr Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 236

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 236

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Glu Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Met Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 237

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 237

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Glu Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Met Val Leu Arg Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 238

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 238

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 239

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 239

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 240

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 240

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 241

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 241

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Thr Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 242

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 242

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 243

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 243

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Thr Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 244

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 244

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Trp Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Thr Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 245

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 245

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 246

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 246

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Thr Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 247

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 247

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Trp Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Leu Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Thr Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 248

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 248

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Arg Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 249

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 249

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 250

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 250

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Arg Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 251

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 251

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Asn Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser Asp Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Arg Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Arg Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 252

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 252

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 253

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 253

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 254

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 254

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 255

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 255

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 256

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 256

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 257

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 257

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 258

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 258

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 259

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 259

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 260

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 260

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 261

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 261

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 262

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 262

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 263

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 263

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 264

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 264

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 265

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 265

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 266

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 266

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 267

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 267

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 268

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 268

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 269

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 269

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 270

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 270

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 271

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 271

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 272

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 272

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 273

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 273

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 274

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 274

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 275

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 275

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 276

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 276

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 277

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 277

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro

325

<210> 278

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 278

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 279

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 279

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 280

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 280

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 281

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 281

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 282

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 282

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 283

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 283

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 284

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 284

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 285

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 285

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 286

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 286

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 287

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 287

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 288

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 288

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 289

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 289

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 290

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 290

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 291

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 291

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 292

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 292

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 293

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 293

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 294

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 294

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 295

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 295

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 296

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 296

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 297

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 297

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 298

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 298

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 299

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 299

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 300

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 300

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 301

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 301

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 302

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 302

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 303

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 303

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 304

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 304

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 305

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 305

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 306

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 306

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 307

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 307

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 308

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 308

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 309

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 309

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 310

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 310

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 311

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 311

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 312

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 312

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 313

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 313

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 314

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 314

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 315

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 315

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 316

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 316

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 317

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 317

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 318

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 318

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 319

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 319

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 320

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 320

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 321

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 321

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 322

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 322

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 323

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 323

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 324

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 324

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 325

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 325

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 326

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 326

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 327

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 327

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 328

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 328

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 329

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 329

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 330

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 330

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 331

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 331

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 332

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 332

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 333

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 333

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 334

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 334

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 335

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 335

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 336

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 336

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 337

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 337

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 338

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 338

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 339

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 339

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 340

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 340

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 341

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 341

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 342

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 342

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 343

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 343

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 344

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 344

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 345

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 345

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 346

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 346

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 347

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 347

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 348

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 348

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 349

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 349

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 350

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 350

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 351

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 351

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 352

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 352

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 353

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 353

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 354

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 354

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 355

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 355

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Thr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 356

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 356

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 357

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 357

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 358

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 358

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 359

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 359

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 360

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 360

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 361

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 361

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 362

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 362

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 363

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 363

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 364

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 364

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 365

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 365

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 366

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 366

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 367

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 367

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 368

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 368

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 369

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 369

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 370

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 370

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 371

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 371

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 372

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 372

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 373

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 373

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 374

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 374

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 375

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 375

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 376

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 376

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 377

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 377

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 378

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 378

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 379

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 379

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 380

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 380

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Arg Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<210> 381

<211> 328

<212> PRT

<213> Artificial sequence

<220>

<223> An artificially synthesized sequence

<400> 381

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Tyr Leu Gly Gly Asp Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Val Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Ile Asp Val Ala His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Pro Val Leu

180 185 190

His Arg Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Lys Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280 285

Leu Tyr Ser Lys Leu Thr Val Lys Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Leu His Glu Ala Leu His Ala His Tyr Thr

305 310 315 320

Arg Lys Glu Leu Ser Leu Ser Pro

325

<---

Claims (108)

1. An antibody that specifically binds to latent myostatin, wherein the antibody comprises (i) HVR-H1 containing the amino acid sequence of SEQ ID NO: 114, HVR-H2 containing the amino acid sequence of SEQ ID NO: 58, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 122, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71 and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (ii) HVR-H1 containing the amino acid sequence of SEQ ID NO: 114, HVR-H2 containing the amino acid sequence of SEQ ID NO: 116, HVR-H3 containing the amino acid sequence of SEQ ID NO: 121, HVR-L1 containing the amino acid sequence of SEQ ID NO: 122, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71 and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (iii) HVR-H1 containing the amino acid sequence of SEQ ID NO: 57, HVR-H2 containing the amino acid sequence of SEQ ID NO: 117, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 122, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71 and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (iv) HVR-H1 containing the amino acid sequence of SEQ ID NO: 57, HVR-H2 containing the amino acid sequence of SEQ ID NO: 118, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 122, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71 and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (v) HVR-H1 containing the amino acid sequence of SEQ ID NO: 57, HVR-H2 containing the amino acid sequence of SEQ ID NO: 119, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 122, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71 and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (vi) HVR-H1 containing the amino acid sequence of SEQ ID NO: 114, HVR-H2 containing the amino acid sequence of SEQ ID NO: 118, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 122, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71, and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (vii) HVR-H1 containing the amino acid sequence of SEQ ID NO: 114, HVR-H2 containing the amino acid sequence of SEQ ID NO: 58, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 123, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71, and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (viii) HVR-H1 containing the amino acid sequence of SEQ ID NO: 115, HVR-H2 containing the amino acid sequence of SEQ ID NO: 58, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 123, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71, and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (ix) HVR-H1 containing the amino acid sequence of SEQ ID NO: 114, HVR-H2 containing the amino acid sequence of SEQ ID NO: 58, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 124, HVR-L2 containing the amino acid sequence of SEQ ID NO: 125, and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74; or (x) HVR-H1 containing the amino acid sequence of SEQ ID NO: 115, HVR-H2 containing the amino acid sequence of SEQ ID NO: 120, HVR-H3 containing the amino acid sequence of SEQ ID NO: 63, HVR-L1 containing the amino acid sequence of SEQ ID NO: 123, HVR-L2 containing the amino acid sequence of SEQ ID NO: 71 and HVR-L3 containing the amino acid sequence of SEQ ID NO: 74. 2. An antibody according to one of subparagraphs 1(i) - (vi) and (ix), containing the framework region of the variable domain of the heavy chain FR1 (H-FR1), containing the amino acid sequence of SEQ ID NO: 132; FR2 heavy chain variable domain framework region (H-FR2), containing the amino acid sequence of SEQ ID NO: 135; FR3 heavy chain variable domain framework region (H-FR3), containing the amino acid sequence of SEQ ID NO: 137; FR4 heavy chain variable domain framework region (H-FR4), containing the amino acid sequence of SEQ ID NO: 138, FR1 light chain variable domain framework (L-FR1) containing the amino acid sequence of SEQ ID NO: 139; a framework region of the FR2 light chain variable domain (L-FR2) containing the amino acid sequence of SEQ ID NO: 140; FR3 light chain variable domain framework (L-FR3) containing the amino acid sequence of SEQ ID NO: 142; And a framework region of the FR4 light chain variable domain (L-FR4) containing the amino acid sequence of SEQ ID NO: 144; or an antibody according to (vii) or (x) of claim 1, containing H-FR1 containing the amino acid sequence of SEQ ID NO: 133; H-FR2 containing the amino acid sequence of SEQ ID NO: 136; H-FR3 containing the amino acid sequence of SEQ ID NO: 137; H-FR4 containing the amino acid sequence of SEQ ID NO: 138; L-FR1 containing the amino acid sequence of SEQ ID NO: 139; L-FR2 containing the amino acid sequence of SEQ ID NO: 140; L-FR3 containing the amino acid sequence of SEQ ID NO: 142; And L-FR4 containing the amino acid sequence of SEQ ID NO: 144; or an antibody according to (vii) or (viii) of claim 1, containing H-FR1 containing the amino acid sequence of SEQ ID NO: 134; H-FR2 containing the amino acid sequence of SEQ ID NO: 136; H-FR3 containing the amino acid sequence of SEQ ID NO: 137; H-FR4 containing the amino acid sequence of SEQ ID NO: 138; L-FR1 containing the amino acid sequence of SEQ ID NO: 139; L-FR2 containing the amino acid sequence of SEQ ID NO: 140; L-FR3 containing the amino acid sequence of SEQ ID NO: 142; And L-FR4 containing the amino acid sequence of SEQ ID NO: 144; or an antibody according to (vii) of claim 1, containing H-FR1 containing the amino acid sequence of SEQ ID NO: 133; H-FR2 containing the amino acid sequence of SEQ ID NO: 136; H-FR3 containing the amino acid sequence of SEQ ID NO: 137; H-FR4 containing the amino acid sequence of SEQ ID NO: 138; L-FR1 containing the amino acid sequence of SEQ ID NO: 139; L-FR2 containing the amino acid sequence of SEQ ID NO: 141; L-FR3 containing the amino acid sequence of SEQ ID NO: 143; and L-FR4 containing the amino acid sequence of SEQ ID NO: 144. 3. An antibody according to claim 1 or 2, characterized in that said antibody comprises: (i) a VH sequence of SEQ ID NO: 86 and a VL sequence of SEQ ID NO: 96; or (ii) the VH sequence of SEQ ID NO: 86 and the VL sequence of SEQ ID NO: 98; or (iii) a VH sequence of SEQ ID NO: 87 and a VL sequence of SEQ ID NO: 96; or (iv) a VH sequence of SEQ ID NO: 88 and a VL sequence of SEQ ID NO: 96; or (v) a VH sequence from SEQ ID NO: 89 and a VL sequence from SEQ ID NO: 96; or (vi) a VH sequence of SEQ ID NO: 90 and a VL sequence of SEQ ID NO: 96; or (vii) a VH sequence of SEQ ID NO: 91 and a VL sequence of SEQ ID NO: 96; or (viii) the VH sequence of SEQ ID NO: 92 and the VL sequence of SEQ ID NO: 97; or (ix) a VH sequence from SEQ ID NO: 92 and a VL sequence from SEQ ID NO: 99; or (x) the VH sequence of SEQ ID NO: 93 and the VL sequence of SEQ ID NO: 97; or (xi) a VH sequence from SEQ ID NO: 94 and a VL sequence from SEQ ID NO: 97; or (xii) a VH sequence from SEQ ID NO: 95 and a VL sequence from SEQ ID NO: 97. 4. An antibody according to one of paragraphs. 1-3, which contains a heavy chain constant region containing the amino acid sequence of SEQ ID NO: 352 and a light chain constant region containing the amino acid sequence of SEQ ID NO: 10. 5. Nucleic acid encoding an antibody according to one of paragraphs. 1-4. 6. The host cell to obtain antibodies according to one of paragraphs. 1-4 containing the nucleic acid according to claim 5. 7. A method for producing an anti-myostatin antibody, comprising culturing a host cell according to claim 6 to obtain an antibody.

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