RU2797348C2 - Antibodies containing only heavy chains that bind to cd22 - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: following is presented: an anti-CD22 antibody containing only heavy chains, a CAR-T molecule that binds to CD22, a multispecific antibody containing a first binding fragment that has a binding specificity for CD22 and a second binding fragment that has a binding affinity for an effector cell. Besides, polynucleotides encoding antibodies, a pharmaceutical composition containing such antibodies, and their use for the treatment of B-cell disorders characterized by CD22 expression are proposed.

EFFECT: invention makes it possible to obtain new antibodies with high affinity for HLA-G.

28 cl, 6 dwg, 4 ex

Description

Cross-reference to related applications

This application claims priority over U.S. Provisional Application No. 62/609,759, filed Dec. 22, 2017, the disclosure of which is hereby incorporated by reference in its entirety.

Technical field

The present invention relates to heavy chain-only human antibodies (eg, ^UniAbs.TM. ) that bind to CD22. This invention also relates to methods for producing such antibodies, compositions, including pharmaceutical compositions, containing such antibodies, and their use in the treatment of B-cell disorders characterized by CD22 expression.

State of the art

CD22

CD22, also known as SIGLEC-2 (UniProt P20273), is a cell surface receptor that is expressed on mature B cells. CD22 contains several Ig domains and is a member of the immunoglobulin superfamily. The extracellular domain of CD22 interacts with sialic acid moieties, including those present on the cell surface protein CD45. CD22 is believed to function as an inhibitory receptor for B cell receptor signaling. Along with CD20 and CD19, the limited expression of CD22 B cells makes it an attractive target for the therapeutic treatment of B cell malignancies. Monoclonal antibodies specific for CD22 have been described in the literature (e.g., Jabbour, Elias, et al. “Monoclonal antibodies in acute lymphoblastic leukemia.” Blood 125.26 (2015): 4010-4016) and have been used therapeutically as standard monoclonal antibody preparations ( e.g. epratuzumab), as well as antibody-drug conjugates (inotuzumab ozogamicin). In addition, anti-CD22 chimeric antigen receptor T cells have been used in clinical practice for the treatment of leukemia (Fry, Terry J., et al. "CD22-targeted CAR T cells induce remission in B-ALL that is naive or resistant to CD19- targeted CAR immunotherapy." Nature medicine (2017)).

Heavy chain antibodies

In a conventional IgG antibody, the association of the heavy chain and the light chain is due in part to a hydrophobic interaction between the light chain constant region and the CH1 constant domain of the heavy chain. There are additional residues in the heavy chain regions of framework 2 (FR2) and framework 4 (FR4) that also contribute to this hydrophobic interaction between the heavy and light chains.

It is known, however, that the serum of camelids (a suborder of Tylopoda that includes camels, dromedaries and llamas) contains a major type of antibody consisting solely of paired H chains (heavy chain-only antibodies or UniAbs™). UniAbs™ Camelidae (Camelus dromedarius, Camelus bactrianus, Lama glama, Lama guanaco, Lama alpaca and Lama vicugna) have a unique structure consisting of one variable domain (VHH), a hinge region and two constant domains (CH2 and CH3) that are highly homologous to the domains CH2 and CH3 of classical antibodies. The ^UniAbs.TM. data do not have a first constant region domain (CH1) that is present in the genome but undergoes splicing during mRNA processing. The absence of the CH1 domain explains the absence of the light chain in UniAbs™, since this domain is the site of anchorage for the light chain constant domain. Such UniAbs™ naturally evolved to confer antigen-binding specificity and high affinity on three CDRs from conventional antibodies or antibody fragments (Muyldermans, 2001; J Biotechnol 74:277-302; Revets et al., 2005; Expert Opin Biol Ther 5:111-124 ). Cartilaginous fish, such as sharks, have also developed a special type of immunoglobulin, designated IgNAR, which lacks polypeptide light chains and is composed entirely of heavy chains. IgNAR molecules can be manipulated by molecular engineering to produce a single heavy chain polypeptide variable domain (vNAR) (Nuttall et al. Eur. J. Biochem. 270, 3543-3554 (2003); Nuttall et al. Function and Bioinformatics 55, 187 -197 (2004); Dooley et al., Molecular Immunology 40, 25-33 (2003)).

The ability of heavy chain-only antibodies lacking a light chain to bind antigen was established in the 1960s (Jaton et al. (1968) Biochemistry, 7, 4185-4195). The immunoglobulin heavy chain, physically separated from the light chain, retained 80% of the antigen-binding activity relative to the tetrameric antibody. Sitia et al. (1990) Cell, 60, 781-790 demonstrated that deletion of the CH1 domain from a rearranged murine µ gene results in a heavy chain-only antibody devoid of a light chain in mammalian cell culture. The generated antibodies retained VH binding specificity and effector functions.

Heavy chain-only antibodies with high specificity and affinity can be generated against various antigens by immunization (van der Linden, R. H., et al. Biochim. Biophys. Acta. 1431, 37-46 (1999)), and the VHH part can be readily cloned and expressed in yeast (Frenken, L. G. J., et al. J. Biotechnol. 78, 11-21 (2000)). Their levels of expression, solubility and stability are significantly higher than those of classical F(ab) or Fv fragments (Ghahroudi, M. A. et al. FEBS Lett. 414, 521-526 (1997)).

Mice in which the λ (lambda) light (L) chain locus and/or the λ and κ (kappa) L chain loci were functionally repressed, and antibodies produced by such mice are described in US Pat. Nos. 7,541,513 and 8,367,888. heavy chain-only antibodies in mice and rats have been described, for example, in WO 2006008548; U.S. Patent Application Publication No. 20100122358; Nguyen et al., 2003, Immunology; 109(1), 93-101; Brüggemann et al., Crit. Rev. Immunol.; 2006, 26(5):377-90; and Zou et al., 2007, J Exp Med; 204(13): 3271–3283. The production of knockout rats by microinjection of zinc finger nuclease into embryos is described in Geurts et al., 2009, Science, 325(5939):433. Soluble heavy chain antibodies and transgenic rodents containing a heterologous heavy chain locus producing such antibodies are described in US Pat. Nos. 8,883,150 and 9,365,655. , in Iri-Sofla et al., 2011, Experimental Cell Research 317:2630-2641 and Jamnani et al., 2014, Biochim Biophys Acta, 1840:378-386.

The essence of the invention

Aspects of the invention relate to antibodies containing only heavy chains, including, without limitation, UniAbs ^TM , with a binding affinity for CD22. Additional aspects of the invention relate to methods for making such antibodies, compositions containing such antibodies, and their use in the treatment of B-cell disorders characterized by CD22 expression.

In some embodiments, a heavy chain-only antibody that binds to CD22 comprises a heavy chain variable region comprising: (a) a CDR1 having two or fewer substitutions in any of the amino acid sequences of SEQ ID NOs: 1-10, and/or ( b) a CDR2 having two or less substitutions in any of the amino acid sequences of SEQ ID NOs: 11-17, and/or (c) a CDR3 having two or less substitutions in any of the amino acid sequences of SEQ ID NOs: 18-23. In some embodiments, the CDR1, CDR2, and CDR3 sequences are present in a human framework. In some embodiments, the heavy chain-only antibody further comprises a heavy chain constant region sequence in the absence of a CH1 sequence.

In some embodiments, the heavy chain-only antibody comprises: (a) a CDR1 sequence selected from the group consisting of SEQ ID NOs: 1-10, and/or (b) a CDR2 sequence selected from the group consisting of SEQ ID NOs: NO: 11-17, and/or (c) a CDR3 sequence selected from the group consisting of SEQ ID NO: 18-23. In some embodiments, the heavy chain-only antibody comprises: (a) a CDR1 sequence selected from the group consisting of SEQ ID NOs: 1-10, and (b) a CDR2 sequence selected from the group consisting of SEQ ID NOs: 11-17, and (c) a CDR3 sequence selected from the group consisting of SEQ ID NOs: 18-23.

In some embodiments, the heavy chain-only antibody comprises: (a) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 11, and a CDR3 sequence of SEQ ID NO: 18, or (b) a CDR1 sequence of SEQ ID NO: 1 , a CDR2 sequence of SEQ ID NO: 12 and a CDR3 sequence of SEQ ID NO: 19, or (c) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 12, and a CDR3 sequence of SEQ ID NO: 20. In some embodiments, the antibody , containing only heavy chains, contains a heavy chain variable region having at least 95% sequence identity with any of the sequences of SEQ ID NO: 24-84. In some embodiments, the heavy chain-only antibody comprises a heavy chain variable region sequence selected from the group consisting of SEQ ID NOs: 24-84. In some embodiments, the heavy chain-only antibody comprises the heavy chain variable region sequence of SEQ ID NO: 24.

In some embodiments, a heavy chain-only antibody that binds to CD22 comprises a heavy chain variable region comprising (a) a CDR1 sequence of the formula:

GX ₁ SIX ₂ X ₃ X ₄ X ₅ X ₆ Y,

where X ₁ represents D or G; X ₂ is S, T, I or N; X ₃ is S or D; X ₄ is G, S or N; X ₅ is D, G or S and X ₆ is Y or H; and (b) a CDR2 sequence of formula:

X ₇ X ₈ YX ₉ GX ₁₀ X ₁₁ ,

where X ₇ represents I or V; X ₈ is Y or H; X ₉ is S or T; X ₁₀ is A, V or S and X ₁₁ is T or A; and (c) a CDR3 sequence of the formula:

X ₁₂ RX ₁₃ DSSX ₁₄ WRS,

where X ₁₂ represents T, A or K; X ₁₃ is D or E and X ₁₄ is N or S.

In some embodiments, a heavy chain-only antibody that binds to CD22 comprises a heavy chain variable region comprising the sequences CDR1, CDR2, and CDR3 in a human VH framework, wherein the CDR sequences are sequences having two or less substitutions in the CDR sequence, selected from the group consisting of SEQ ID NO: 1-23.

In some embodiments, the heavy chain-only antibody comprises a heavy chain variable region comprising the sequences CDR1, CDR2, and CDR3 in a human VH framework, the CDR sequences being selected from the group consisting of SEQ ID NOs: 1-23.

In some embodiments, a heavy chain-only antibody that binds to CD22 comprises a heavy chain variable region comprising: (a) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 11, and a CDR3 sequence of SEQ ID NO: 18, or (b) CDR1 SEQ ID NO: 1, CDR2 SEQ ID NO: 12, and CDR3 SEQ ID NO: 19, or (c) CDR1 SEQ ID NO: 1, CDR2 SEQ ID NO: 12, and CDR3 SEQ ID NO: 20 in human VH framework.

In some embodiments, the heavy chain-only antibody is multispecific. In some embodiments, the heavy chain-only antibody is bispecific. In some embodiments, the heavy chain-only antibody has binding affinity for two different CD22 proteins. In some embodiments, the heavy chain-only antibody has binding affinity for two different epitopes on the same CD22 protein. In some embodiments, the heavy chain-only antibody has binding affinity for an effector cell. In some embodiments, the heavy chain-only antibody has binding affinity for a T cell antigen. In some embodiments, the heavy chain-only antibody has a binding affinity for CD3. In some embodiments, the heavy chain-only antibody is in CAR-T format.

Aspects of the invention relate to pharmaceutical compositions containing a heavy chain-only antibody as described herein.

Aspects of the invention relate to methods for treating a CD22-expressing B-cell disorder comprising administering to a subject with said disorder an antibody or pharmaceutical composition as described herein. In certain other aspects, the invention relates to the use of an antibody described herein in the manufacture of a medicament for the treatment of a B cell disorder characterized by CD22 expression. In yet other aspects, the invention provides an antibody described herein for use in the treatment of a B cell disorder characterized by CD22 expression. With respect to these aspects, and in some embodiments, the disorder is diffuse large B-cell lymphoma (DLBCL). In some embodiments, the disorder is non-Hodgkin's lymphoma (NHL). In some embodiments, the disorder is systemic lupus erythematosus (SLE). In some embodiments, the disorder is rheumatoid arthritis (RA). In some embodiments, the disorder is multiple sclerosis (MS).

Aspects of the invention relate to polynucleotides encoding the antibody described herein, vectors containing such polynucleotides, and cells containing such vectors.

Aspects of the invention relate to methods for producing an antibody described herein, including growing a cell described herein under conditions favorable for expression of the antibody, and isolating the antibody from the cell.

Aspects of the invention relate to methods for producing an antibody described herein, including immunizing a UniRat animal with CD22 and identifying heavy chain CD22 binding sequences.

These and other aspects will be further explained in the rest of the disclosure, including examples.

Brief description of graphic materials

On FIG. 1 shows the unique amino acid sequences of the CDRs of an anti-CD22 heavy chain antibody.

On FIG. 2 shows the amino acid sequences of an anti-CD22 antibody heavy chain variable domain.

On FIG. 3 shows the amino acid sequences of CDR1, CDR2 and CDR3 of an anti-CD22 heavy chain antibody.

On FIG. 4 shows the biological activity of an anti-CD22 antibody containing heavy chains only.

On FIG. 5A is a graph showing percent specific lysis as a function of antibody concentration for Daudi cells.

On FIG. 5B is a graph showing percent specific lysis as a function of antibody concentration for Raji cells.

On FIG. 5C is a graph showing percent specific lysis as a function of antibody concentration for Ramos cells.

On FIG. 5D is a schematic illustration of an anti-CD22xCD3 bispecific antibody in accordance with one embodiment of the invention.

On FIG. 6 is a series of graphs showing serum titer as a function of dilution.

Detailed Description of the Preferred Embodiments

In the practice of the present invention, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are known to those skilled in the art, are used. Such methods are described in detail in the literature, for example, "Molecular Cloning: A Laboratory Manual", second edition (Sambrook et al., 1989); "Oligonucleotide Synthesis" (M. J. Gait, ed., 1984); "Animal Cell Culture" (R. I. Freshney, ed., 1987); "Methods in Enzymology" (Academic Press, Inc.); “Current Protocols in Molecular Biology” (F. M. Ausubel et al., eds., 1987, and periodic updates); “PCR: The Polymerase Chain Reaction”, (Mullis et al., ed., 1994); “A Practical Guide to Molecular Cloning” (Perbal Bernard V., 1988); “Phage Display: A Laboratory Manual” (Barbas et al., 2001); Harlow, Lane and Harlow, Using Antibodies: A Laboratory Manual: Portable Protocol No. I, Cold Spring Harbor Laboratory (1998); and Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory; (1988).

When a range of values is given, it is to be understood that this invention covers every intermediate value, to the tenth decimal place of the lower limit, unless the context expressly states otherwise, between the upper and lower limits of that range, as well as any other specified or intermediate value within such specified range. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges and are also included in this invention, subject to any specifically excluded limit in the specified range. If the specified range includes one or both of the limits, ranges excluding one or both of these included limits are also included in this invention.

Unless otherwise indicated, antibody residues herein are numbered according to the Kabat numbering system (eg, Kabat et al., Sequences of Immunological Interest. ^5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991) ).

In order to provide a thorough understanding of the present invention, numerous specific details are set forth in the following detailed description. However, for a person skilled in the art it will be obvious that the present invention can be practiced without one or more of these specific details. In other cases, well-known features and procedures well known to those skilled in the art have not been described in order to avoid obscuring the invention.

All references herein, including patent applications and publications, are incorporated herein by reference in their entirety.

I. Definitions

The term "comprising" means that the listed elements are required for the composition/method/kit, however, to form the composition/method/kit, etc. other elements may be included within the scope of the claims.

The term "consisting essentially of" means limiting the scope of the described composition or method to the specified materials or step-by-step actions that do not significantly affect the main and new characteristic (s) of the subject matter of the invention.

By "consisting of" is meant the exclusion of any element, step, or ingredient not listed in a claim from a composition, method, or kit.

Antibody residues herein are numbered according to the Kabat numbering system and the EU numbering system. The Kabat numbering system is commonly used to designate a residue in a variable domain (approximately heavy chain residues 1-113) (e.g., Kabat et al., Sequences of Immunological Interest. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. ( 1991)). The "EU numbering system" or "EU index" is generally used to refer to a residue in an immunoglobulin heavy chain constant region (eg, the EU index described in Kabat et al., supra). "EU index according to Kabat" refers to the EU residue numbering of a human IgG1 antibody. Unless otherwise indicated, references herein to residue numbers in the variable domain of antibodies mean the numbering of residues in accordance with the Kabat numbering system. Unless otherwise indicated, references herein to residue numbers in the constant domain of antibodies mean the numbering of residues in accordance with the EU numbering system.

The term "monoclonal antibody" as used herein refers to an antibody derived from a population of substantially homogeneous antibodies, i.e. individual antibodies within the population are identical except for naturally occurring mutations that may be present in small amounts. Monoclonal antibodies are highly specific and directed against a single antigenic site. In addition, unlike conventional (polyclonal) antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on an antigen. Monoclonal antibodies in accordance with this invention can be obtained by the hybridoma method, first described by Kohler et al. (1975) Nature 256:495 can also be obtained, for example, by methods for producing a recombinant protein (see, for example, US patent No. 4816567).

The term "variable" as used herein in connection with antibodies refers to the fact that the sequences of certain portions of the variable domains of an antibody vary greatly between antibodies and contribute to the binding and specificity of each particular antibody for its particular antigen. At the same time, the variability is not evenly distributed across the variable domains of antibodies. It is concentrated in three segments of both the light and heavy chain variable domains called hypervariable regions. More conserved fragments of variable domains are called framework regions (FRs). Variable domains of native light and heavy chains contain four FRs, predominantly taking on the β-sheet configuration and connected by three hypervariable regions forming loops connecting β-type structures, and in some cases being part of them. The hypervariable regions of each chain are held together in close proximity to the FR and, together with the hypervariable regions of the other chain, are involved in the formation of the antibody antigen-binding site (see Kabat et al., Sequences of Proteins of Immunological Interest, ^5th Ed. Public Health Service, National Institutes of Health, Bethesda, M.D. (1991)). The constant domains are not directly involved in the binding of the antibody to the antigen, but exhibit various effector functions, for example, participation of the antibody in antibody-dependent cell-mediated cytotoxicity (ADCC).

The term "hypervariable region" as used herein refers to the amino acid residues of an antibody that are responsible for antigen binding. The hypervariable region typically contains amino acid residues from the "complementarity determining region" or "CDR" (e.g., residues 31-35 (H1), 50-65 (H2) and 95-102 (H3) in the heavy chain variable domain; Kabat et al ., Sequences of Proteins of Immunological Interest, ^5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and/or residues from hypervariable loop residues 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). "Framework region" or "FR" residues, as defined herein, are hypervariable region residues other than variable domain residues.

Exemplary CDR designations are shown herein, however, one skilled in the art will appreciate that a number of CDR definitions are commonly used, including the Kabat definition (see “Zhao et al. A germline knowledge based computational approach for determining antibody complementarity determining regions.” Mol Immunol. 2010;47:694–700), which is based on sequence variability and is the most commonly used. The Chothia definition is based on the location of regions of closed structure (Chothia et al. “Conformations of immunoglobulin hypervariable regions.” Nature. 1989; 342:877–883). Alternative definitions of CDR of interest include, without limitation, those disclosed by Honegger, “Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool.” J Mol Biol. 2001;309:657–670; Ofran et al. “Automated identification of complementarity determining regions (CDRs) reveals peculiar characteristics of CDRs and B cell epitopes.” J Immunol. 2008;181:6230–6235; Almagro “Identification of differences in the specificity-determining residues of antibodies that recognize antigens of different size: implications for the rational design of antibody repertoires.” J Mol Recognit. 2004;17:132–143; and Padlanet al. “Identification of specificity-determining residues in antibodies.” Faseb J. 1995;9:133–139., the contents of each of which are incorporated herein in their entirety by reference.

The terms "antibody containing only heavy chains", "antibody with heavy chains" are used interchangeably herein and refer in the broadest sense to antibodies that lack the light chain of a conventional antibody. The terms specifically include, without limitation, homodimeric antibodies containing a VH antigen-binding domain and CH2 and CH3 constant domains, in the absence of a CH1 domain; functional (antigen-binding) variants of such antibodies, soluble VH, Ig-NAR variants containing a homodimer of one variable domain (V-NAR) and five C-like constant domains (C-NAR), and functional fragments thereof; and soluble single domain antibodies (sUniDabs™). In one embodiment, a heavy chain-only antibody consists of a variable region antigen-binding domain consisting of framework 1, CDR1, framework 2, CDR2, framework 3, CDR3, and framework 4. In another embodiment, the heavy chain-only antibody consists of from an antigen-binding domain of at least a portion of the hinge region and the CH2 and CH3 domains. In another embodiment, the heavy chain-only antibody consists of an antigen-binding domain of at least a portion of the hinge region and a CH2 domain. In a further embodiment, the heavy chain-only antibody consists of an antigen-binding domain, at least a portion of the hinge region, and a CH3 domain. Heavy chain-only antibodies in which the CH2 and/or CH3 domain is truncated are also included in this document. In a further embodiment, the heavy chain consists of an antigen-binding domain and at least one CH domain (CH1, CH2, CH3, or CH4), but without the hinge region. The heavy chain-only antibody may be in the form of a dimer in which two heavy chains are disulfide bonded to each other, covalently or non-covalently bonded to each other. An antibody containing only heavy chains may belong to the IgG subclass, but antibodies belonging to other subclasses such as the IgM, IgA, IgD, and IgE subclass are also included herein. In a specific embodiment, the heavy chain-only antibody has an IgG1, IgG2, IgG3, or IgG4 subtype, in particular an IgG1 subtype. In one embodiment, heavy chain-only antibodies are used herein as the binding (targeting) domain of a chimeric antigen receptor (CAR). The definition specifically includes human heavy chain-only antibodies called ^UniAbs.TM . produced by transgenic rats with human immunoglobulin ( ^UniRat.TM .). The variable regions (VH) of UniAbs ^TM are referred to as UniDabs ^TM , and are versatile building blocks that can be linked to Fc regions or serum albumin to develop new therapeutics with multispecificity, increased potency and extended half-life. Because homodimeric ^UniAbs.TM. lack the light chain and hence the VL domain, the antigen is recognized by a single domain, ie the heavy chain variable domain of the heavy chain-only (VH) antibody.

The terms "CD22" and "cluster of differentiation-22" as used herein refer to the lectin molecule of the SIGLEC family found on the surface of mature B cells and to a lesser extent in some immature B cells. The term "CD22" includes human CD22 protein and any non-human animal species, and specifically includes human CD22 as well as non-human mammalian CD22.

As used herein, the term "human CD22" includes any variants, isoforms, and species homologues of human CD22 (UniProt P20273), regardless of their source or method of preparation. Thus, "human CD22" includes human CD22 naturally expressed by cells and CD22 expressed in cells transfected with the human CD22 gene.

The terms "anti-CD22 heavy chain-only antibody", "anti-CD22 heavy chain-only antibody", "anti-CD22 heavy chain-only antibody", and "anti-CD22 heavy chain-only antibody" are used interchangeably herein. to denote an antibody containing only heavy chains, as defined above, for immunospecific binding to CD22, including human CD22, as defined above. The definition includes, without limitation, heavy chain-only human antibodies produced by transgenic animals such as transgenic rats or transgenic mice expressing human immunoglobulin, including UniRats ^™ producing anti-human CD22 UniAb™ antibodies as defined above.

"Percent (%) Amino Acid Sequence Identity" in relation to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in a reference polypeptide sequence, after alignment of the sequences and insertion of gaps, if necessary, to achieve the maximum percent sequence identity, and not including any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be performed in a variety of ways known to one of skill in the art, for example using open source software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR). 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. For the purposes of this document, however, % amino acid sequence identity values are obtained using the ALIGN-2 sequence comparison program.

An "isolated" antibody is one that has been identified and separated and/or recovered from a component of its natural environment. The contaminant component of the natural environment is substances that interfere with the diagnostic or therapeutic use of the antibody and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In preferred embodiments, the antibody will be purified to (1) greater than 95% by weight of the antibody as determined by Lowry's method, and most preferably greater than 99% by weight, (2) sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a rotating beaker sequencer, or (3) to homogeneity (SDS-PAGE) under reducing or non-reducing conditions using Coomassie blue stain or, preferably, a silver-based stain. The isolated antibody includes the antibody in situ in recombinant cells, since at least one component of the natural environment of the antibody will be absent. Typically, however, isolated antibody will be obtained by at least one purification step.

The antibodies of the present invention include multispecific antibodies. Multispecific antibodies have more than one binding specificity. The term "multispecific" specifically includes "bispecific" and "trispecific", as well as higher order independent specific binding affinity, such as higher order polyepitope specificity, as well as tetravalent antibodies and antibody fragments. "Multispecific" antibodies specifically include antibodies containing a combination of different binding entities, as well as antibodies containing more than one and the same binding entity. The terms “multispecific antibody”, “heavy chain only multispecific antibody”, “heavy chain multispecific antibody”, and “multispecific UniAb™” are used herein in their broadest sense and encompass all antibodies with more than one binding specificity. The multispecific anti-CD22 heavy chain antibodies of the present invention specifically include antibodies that immunospecifically bind to more than one non-overlapping epitope on the CD22 protein, such as human CD22.

An "epitope" is a region on the surface of an antigen molecule to which one antibody molecule binds. Typically, an antigen has several or many different epitopes and reacts with many different antibodies. The term particularly includes linear epitopes and conformational epitopes.

"Epitope mapping" is the process of identifying antibody binding sites or epitopes on their target antigens. Antibody epitopes may be linear or conformational epitopes. Linear epitopes are formed by a continuous sequence of amino acids in a protein. Conformational epitopes are formed from amino acids that are discontinuous in the sequence of a protein, but which come together when the protein folds into its three-dimensional structure.

The term "polyepitope specificity" refers to the ability to specifically bind to two or more different epitopes on the same or different targets. As noted above, the invention specifically includes anti-CD22 heavy chain-only antibodies with polyepitope specificity, i.e., anti-CD22 heavy chain-only antibodies that bind to two or more non-overlapping epitopes on a CD22 protein, such as human CD22. . The term "non-overlapping epitope(s)" or "non-competitive epitope(s)" of an antigen is defined herein as meaning an epitope(s) that are recognized by one member of an antigen-specific antibody pair but not by another member. Pairs of antibodies or antigen-binding regions that target the same antigen on a multispecific antibody that recognize non-overlapping epitopes do not compete for binding to that antigen and are able to bind that antigen simultaneously.

An antibody binds "substantially the same epitope" as a reference antibody when the two antibodies recognize identical or sterically overlapping epitopes. The most widely used and rapid methods for determining whether two epitopes bind to identical or sterically overlapping epitopes are competitive binding assays, which can be configured in a variety of different formats using either labeled antigen or labeled antibody. Typically, the antigen is immobilized in a 96-well plate and the ability of unlabeled antibodies to block the binding of labeled antibodies is measured using radioactive or enzyme labels.

The term "valent", in the context of this document, refers to the specified number of binding sites in the antibody molecule.

A "polyvalent" antibody has two or more binding sites. Thus, the terms "bivalent", "trivalent" and "tetravalent" refer to the presence of two binding sites, three binding sites and four binding sites, respectively. Thus, a bispecific antibody of the invention is at least divalent and may be trivalent, tetravalent, or otherwise polyvalent.

A wide variety of protein methods and configurations are known and are used to generate bispecific monoclonal antibodies (BsMAB), trispecific antibodies, and the like.

The term "bispecific three-chain antibody-like molecule" or "TCA" is used herein to refer to antibody-like molecules comprising, consisting essentially of, or consisting of three polypeptide subunits, two of which contain, consist essentially of, or consist of one heavy and one light chain a monoclonal antibody, or functional antigen-binding fragments of such antibody chains, comprising an antigen-binding region and at least one CH domain. This heavy chain/light chain pair has binding specificity for the first antigen. The third polypeptide subunit contains, consists essentially of, or consists of a heavy chain-only antibody containing an Fc region containing CH2 and/or CH3 and/or CH4 domains, in the absence of a CH1 domain, and an antigen-binding domain that binds an epitope of a second antigen or another epitope of the first antigen, wherein such binding domain is derived from or has sequence identity to the heavy or light chain variable region of the antibody. Portions of such a variable region may be encoded by V _H and/or V _L gene segments, D and J _H gene segments, or J _L gene segments. The variable region may be encoded by rearranged V _H DJ _H , V _L DJ _H , V _H J _L or V _L J _L gene segments. The TCA protein uses a heavy chain-only antibody as defined above.

The term "chimeric antigen receptor" or "CAR" is used herein in its broadest sense to refer to an engineered receptor that grafts a desired binding specificity (e.g., the antigen-binding region of a monoclonal antibody or other ligand) to the membrane-spanning and intracellular signaling domains. Typically, the receptor is used to graft the specificity of a monoclonal antibody onto a T cell to create chimeric antigen receptors (CARs). (J Natl Cancer Inst, 2015; 108(7):dvj439; and Jackson et al., Nature Reviews Clinical Oncology, 2016; 13:370–383.)

The term "human antibody" is used herein to include antibodies that have variable and constant regions derived from human germline immunoglobulin sequences. As used herein, human antibodies may include amino acid residues not encoded by germline human immunoglobulin sequences, such as mutations introduced by in vitro random or site-specific mutagenesis or by in vivo somatic mutation. The term "human antibody" specifically includes heavy chain-only antibodies having human heavy chain variable region sequences produced by transgenic animals such as transgenic rats or mice, in particular UniAbs™ produced by UniRats™ as defined above.

By "chimeric antibody" or "chimeric immunoglobulin" is meant an immunoglobulin molecule containing amino acid sequences from at least two different Ig loci, for example, a transgenic antibody containing a portion encoded by a human Ig locus and a portion encoded by a rat Ig locus. Chimeric antibodies include transgenic antibodies with non-human Fc regions or artificial Fc regions, and human idiotypes. Such immunoglobulins may be isolated from animals of the invention that have been engineered to produce such chimeric antibodies.

As used herein, the term "effector cell" refers to an immune cell that participates in the effector phase of the immune response, as opposed to the cognitive and activation phases of the immune response. Some effector cells express specific Fc receptors and perform specific immune functions. In some embodiments, an effector cell, such as a natural killer cell, is capable of inducing antibody-dependent cellular cytotoxicity (ADCC). For example, monocytes and macrophages that express FcR are involved in the specific killing of target cells and the presentation of antigens to other components of the immune system or bind to cells that present antigens. In some embodiments, the effector cell may phagocytose the target antigen or target cell.

"Human effector cells" are leukocytes that express receptors such as T cell receptors or FcRs and perform effector functions. Preferably, such cells express at least FcγRIII and perform an ADCC effector function. Examples of human leukocytes that mediate ADCC include natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils; wherein NK cells are preferred. Effector cells can be isolated from a native source, such as blood or PBMCs, as described herein.

The term "immune cell" is used herein in its broadest sense, including, without limitation, cells of myeloid or lymphoid origin, such as lymphocytes (such as B cells and T cells, including cytolytic T cells (CTLs)), killer cells, natural killer (NK) cells, macrophages, monocytes, eosinophils, polymorphonuclear cells such as neutrophils, granulocytes, mast cells and basophils.

The "effector functions" of an antibody refer to that biological activity that is related to the Fc region (native sequence Fc region or amino acid sequence variant Fc region) of the antibody. Examples of antibody effector functions include C1q binding; complement-dependent cytotoxicity; binding to the Fc receptor; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; suppression of cell surface receptors. (e.g. B cell receptor, BCR), etc.

"Antibody-dependent cell-mediated cytotoxicity" and "ADCC" refer to a cell-mediated response in which non-specific cytotoxic cells that express Fc receptors (FcRs) (e.g., natural killer (NK) cells, neutrophils, and macrophages) recognize the bound antibody to the target cell and thus lead to lysis of the target cell. The primary cells mediating ADCC, natural killer cells, only express FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. FcR expression data on hematopoietic cells are summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). The in vitro ADCC assay described in US Pat. No. 5,500,362 or US Pat. No. 5,821,337 can be performed to assess ADCC activity of a molecule of interest. Effector cells suitable for such an assay include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively or in addition, the ADCC activity of the molecule of interest can be assessed in vivo, eg in an animal model, eg as described in Clynes et al. PNAS (USA) 95:652-656 (1998).

"Complement dependent cytotoxicity" or "CDC" refers to the ability of a molecule to lyse a target in the presence of complement. The complement activation pathway is initiated by the binding of the first component of the complement system (C1q) to a molecule (eg, antibody) that forms a complex with a recognized antigen. To evaluate complement activation, a CDC assay can be performed, for example, as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996).

"Binding affinity" refers to the strength of the total number of non-covalent interactions between a single binding site of a molecule (eg, an antibody) and its binding partner (eg, an antigen). Unless otherwise indicated, as used herein, "binding affinity" refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (eg, antibody and antigen). The affinity of a molecule X for its partner Y as a whole can be expressed by the dissociation constant (Kd). Affinity can be measured by conventional methods known in the art. Low affinity antibodies usually bind antigen slowly and tend to dissociate easily, while high affinity antibodies usually bind antigen faster and tend to remain bound.

As used herein, the term "Kd" or "Kd value" refers to the dissociation constant determined by BioLayer interferometry using an Octet QK384 instrument (Fortebio Inc., Menlo Park, CA) in kinetic mode. For example, anti-mouse Fc sensors are loaded with mouse Fc antigen fused and then loaded into wells containing antibodies to measure concentration-dependent rates (kon). Antibody dissociation rates (koff) are measured in the last step when the probes are immersed in wells containing only buffer. Kd is the koff/kon ratio. (For details, see Concepcion, J, et al., Comb Chem High Throughput Screen, 12(8), 791-800, 2009).

The terms "treatment", "treat" and the like are used herein to denote the achievement of the desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of completely or partially preventing a disease or its symptoms and/or may be therapeutic in terms of partially or completely curing a disease and/or an adverse event associated with the disease. As used herein, the term "treatment" encompasses any treatment of a disease in a mammal, and includes: (a) preventing the onset of a disease in a subject who may be predisposed to the disease but who has not yet been diagnosed with the disease; (b) inhibition of the disease, ie. cessation of its development; or (c) alleviating the disease, i. e. disease regression. The therapeutic agent may be administered before, during, or after the onset of a disease or injury. The treatment of the current disease, in which the treatment stabilizes or reduces the undesirable clinical symptoms of the patient, is of particular interest. Such treatment is desirable to carry out until the complete loss of function in the affected tissues. Therapy may be administered to a subject during the symptomatic stage of the disease, and in some cases after the symptomatic stage of the disease.

The term "therapeutically effective amount" means the amount of an active agent that is necessary to confer a therapeutic effect in a subject. For example, a "therapeutically effective amount" is an amount that causes, ameliorates, or otherwise causes improvement in pathological symptoms, disease progression, or physiological conditions associated with the disease, or which enhances resistance to a disorder.

The terms "B-cell neoplasms" or "mature B-cell neoplasms" in the context of this invention include small cell lymphocytic lymphoma, B-cell prolymphocytic lymphoma, B-cell chronic lymphocytic leukemia, mantle cell lymphoma, Burkitt's lymphoma, follicular lymphoma, diffuse B Large cell lymphoma (DLBCL), multiple myeloma, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, plasma cell neoplasms such as plasma cell myeloma, plasmacytoma, monoclonal immunoglobulin deposition disease, heavy chain disease, MALT lymphoma, nodular B-cell lymphoma marginal zone, intravascular large B-cell lymphoma, primary effusion lymphoma, lymphomatoid granulomatosis, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, primary effusion lymphoma and AIDS-associated non-Hodgkin's lymphoma.

The terms "subject", "individual" and "patient" are used interchangeably herein to refer to a mammal that is being evaluated for and/or being treated. In one embodiment, the mammal is a human. The terms "subject", "individual" and "patient" include, without limitation, individuals with cancer, individuals with autoimmune diseases, pathogenic infections, and the like. The subjects may be humans, but may also be other mammals, in particular those mammals that can be used as laboratory models of human disease, such as mice, rats, etc.

The term "pharmaceutical composition" refers to a formulation that is in a form that allows the biological activity of the active ingredient to be effective and that does not contain additional components that are unacceptably toxic to the subject to whom the composition will be administered. Such formulations are sterile. "Pharmaceutical excipients" (carriers, adjuvants) are those that can reasonably be administered to a mammalian subject and provide an effective dose of the active ingredient used.

A "sterile" composition is aseptic or free from, or substantially free of, any living microorganisms and their spores. "Frozen" composition is a composition at a temperature below 0ºC.

A "stable composition" is one in which the protein substantially retains its physical stability and/or chemical stability and/or biological activity during storage. Preferably, the composition substantially retains its physical or chemical stability as well as its biological activity. The storage period is generally selected based on the shelf life of the composition. A variety of techniques for measuring protein stability are known in the art and are discussed in Peptide and Protein Drug Delivery, 247-301. Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones. A. Adv. Drug Delivery Rev. 10:29-90) (1993), for example. Stability can be measured at a selected temperature for a selected period of time. Stability can be assessed qualitatively and/or quantitatively in a variety of ways, including assessing the formation of aggregates (for example, using size exclusion chromatography, by measuring turbidity and/or visual control); by evaluating charge heterogeneity using cation exchange chromatography, isoelectric image focusing (icIEF), or capillary electrophoresis; amino-terminal or carboxy-terminal sequence analysis; mass spectrometric analysis; SDS-PAGE analysis to compare recovered and intact antibodies; peptide map analysis (eg tryptic or LYS-C); assessing the biological activity or antigen-binding function of the antibody; and so on. The instability may include one or more of: aggregation, deamidation (e.g. Asn deamidation), oxidation (e.g. Met oxidation), isomerization (e.g. Asp isomerization), clipping/hydrolysis/fragmentation (e.g. hinge fragmentation), formation succinimide, unpaired cysteine(s), N-terminal extension, C-terminal processing, glycosylation differences, and the like.

II. Detailed Description of the Invention

Antibodies to CD22

The present invention relates to a family of closely related heavy chain-only antibodies that bind to human CD22. The antibodies of this family contain the CDR sequences as defined herein and shown in FIG. 1 and are illustrated by the heavy chain variable region (VH) sequences of SEQ ID NOs: 24-84 shown in FIG. 2. Families of antibodies provide a number of advantages that favor their use as clinical(s) therapeutic(s) agent(s). Antibodies include members with a range of binding affinities allowing the selection of a particular sequence with the desired binding affinity.

A suitable antibody may be selected from those provided herein for development and therapeutic or other uses, including, but not limited to, use as a bispecific antibody such as that shown in FIG. 5B, or a trispecific antibody or part of a CAR-T structure.

Determination of affinity for a candidate protein can be performed using methods known in the art, such as Biacore measurements. Members of the antibody family may have an affinity for CD22 with Kd from about 10 ^-6 to about 10 ^-11 , including, but not limited to: from about 10 ^-6 to about 10 ^-10 ; from about 10 ^-6 to about 10 ^-9 ; from about 10 ^-6 to about 10 ^-8 ; from about 10 ^-8 to about 10 ^-11 ; about 10 ^-8 to about 10 ^-10 ; from about 10 ^-8 to about 10 ^-9 ; from about 10 ^-9 to about 10 ^-11 ; from about 10 ^-9 to about 10 ^-10 ; or any value in those ranges. The choice of affinity can be confirmed by a biological assessment for modulation, eg blocking, CD22 biological activity, including in vitro assays, preclinical models and clinical trials, and an assessment of potential toxicity.

The antibody family members described herein are not cross-reactive with the Cynomolgus CD22 protein, but can be designed to be cross-reactive with the Cynomolgus CD22 protein or CD22 proteins of other species, as appropriate.

The CD22-specific antibody family herein includes a VH domain containing the sequences CDR1, CDR2, and CDR3 in a human VH framework. CDR sequences can be located, for example, in the region around amino acid residues 26-35; 53-59; and 98-117, for CDR1, CDR2, and CDR3, respectively, provided exemplary variable region sequences set forth in SEQ ID NOs: 24-84. One of ordinary skill in the art will appreciate that the CDR sequences may be in different positions if a different framework sequence is chosen, although the order of the sequences generally remains the same.

The CDR1, CDR2 and CDR3 sequences of the anti-CD22 antibodies of this invention may be encompassed by the following structural formulas, wherein X is a variable amino acid, which may be specific amino acids as indicated below.

CDR1

GX ₁ SIX ₂ X ₃ X ₄ X ₅ X ₆ Y,

where X ₁ represents D or G;

X ₂ is S, T, I or N;

X ₃ is S or D;

X ₄ is G, S, or N;

X ₅ is D, G or S and

X ₆ is Y or H.

CDR2

X ₇ X ₈ YX ₉ GX ₁₀ X ₁₁ ,

where X ₇ represents I or V;

X ₈ is Y or H;

X ₉ is S or T;

X ₁₀ is A, V or S and

X ₁₁ is T or A.

CDR3

X ₁₂ RX ₁₃ DSSX ₁₄ WRS,

where X ₁₂ represents T, A or K;

X ₁₃ is D or E and

X ₁₄ is N or S.

Exemplary CDR1, CDR2, and CDR3 sequences are shown in FIG. 1 and FIG. 3.

In some embodiments, the heavy chain-only anti-CD22 antibody of the invention comprises a CDR1 sequence of any one of SEQ ID NOs: 1-10. In a specific embodiment, the CDR1 sequence is SEQ ID NO: 1.

In some embodiments, the heavy chain-only anti-CD22 antibody of the invention comprises a CDR2 sequence of any one of SEQ ID NOs: 11-17. In a specific embodiment, the CDR2 sequence is SEQ ID NO: 11.

In some embodiments, the heavy chain-only anti-CD22 antibody of the invention comprises a CDR3 sequence of any of SEQ ID NOs: 18-23. In a specific embodiment, the CDR2 sequence is SEQ ID NO: 18.

In a further embodiment, the heavy chain-only anti-CD22 antibody of the present invention comprises the sequence CDR1 of SEQ ID NO:1; a CDR2 sequence of SEQ ID NO: 11; and a CDR3 sequence of SEQ ID NO: 18.

In further embodiments, the heavy chain-only CD22 antibody of the invention comprises any of the heavy chain variable region amino acid sequences of SEQ ID NOs: 24 to 84 (FIG. 2).

In another further embodiment, the heavy chain-only CD22 antibody of the present invention comprises the heavy chain variable region of SEQ ID NO: 24.

In some embodiments, the CDR sequence in an anti-CD22 heavy chain-only antibody of the invention contains one or two amino acid substitutions relative to a CDR1, CDR2 and/or CDR3 sequence or a set of CDR1, CDR2 and CDR3 sequences in any of SEQ ID NOs: 1-23 (Fig. 1). In some embodiments, said amino acid substitution(s) is one or two CDR1 amino acid positions 4-6 and/or one or two CDR2 amino acid positions 2, 4-7, and/or one or two CDR3 amino acid positions 5-12, relative to the formulas above. In some embodiments, heavy chain-only CD22 antibodies will comprise a heavy chain variable region sequence with at least about 85% identity, at least 90% identity, at least 95% identity, at least 98% identity, or at least 99% identity to any of the heavy chain variable region sequences of SEQ ID NOs: 24-84 (shown in FIG. 2).

In some embodiments, bispecific or multispecific antibodies are provided, which may have any of the configurations discussed herein, including, without limitation, a bispecific three-chain antibody-like molecule. In some embodiments, the bispecific antibody may comprise at least one heavy chain variable region having a binding specificity for CD22 and at least one heavy chain variable region having a binding specificity for a protein other than CD22. In some embodiments, the bispecific antibody may comprise a heavy chain/light chain pair that has binding specificity for the first antigen, and a heavy chain from a heavy chain-only antibody containing an Fc portion containing CH2 and/or CH3 and/or CH4 domains, in the absence of a CH1 domain, and an antigen-binding domain that binds an epitope of the second antigen or another epitope of the first antigen. In one particular embodiment, the bispecific antibody comprises a heavy chain/light chain pair that has binding specificity for an antigen on the effector cell (e.g., CD3 protein on a T cell), and the heavy chain from the heavy chain-only antibody contains an antigen-binding domain, having binding specificity for CD22.

In some embodiments, when the protein of the invention is a bispecific antibody, one arm of the antibody (one binding fragment) is specific for human CD22 while the other fragment may be specific for target cells, tumor-associated antigens, target antigens, for example, integrins, etc., pathogen antigens, checkpoint proteins, and the like. Target cells specifically include cancer cells, including, without limitation, cells from a hematological tumor, such as B cell tumors, as described above.

Various formats of bispecific antibodies are within the scope of the invention, including, without limitation, single chain polypeptides, double chain polypeptides, triple chain polypeptides, four chain polypeptides, and polypeptide multiples thereof. As used herein, bispecific antibodies specifically include T cell bispecific antibodies that bind to CD22, which is selectively expressed on mature B cells, and CD3 (antibodies to CD22xCD3). Such antibodies induce strong T-cell mediated killing of cells expressing CD22.

Production of anti-CD22 antibodies containing only heavy chains

The heavy chain-only antibodies of this invention can be prepared using methods known in the art. In a preferred embodiment, the heavy chain-only antibodies are here produced by transgenic animals, including transgenic mice and rats, preferably rats, in which endogenous immunoglobulin genes are knocked out or disabled. In a preferred embodiment, the heavy chain-only antibodies of the present invention are produced in UniRat™. UniRat™ have silent endogenous immunoglobulin genes and use the human immunoglobulin heavy chain translocus to express a diverse, naturally optimized repertoire of fully human HCAbs. While endogenous immunoglobulin loci in rats can be knocked out or silenced using a variety of technologies, in UniRat™, zinc finger (endo)nuclease (ZNF) technology has been used to inactivate the endogenous rat heavy chain J locus, light chain Cκ locus, and the Cλ locus of the light chain. ZNF constructs for microinjection into oocytes can produce knockout (KO) lines of IgH and IgL. For details, see, for example, Geurts et al., 2009, Science 325:433. Characterization of Ig heavy chain knockout rats has been reported by Menoret et al., 2010, Eur. J. Immunol. 40:2932-2941. The advantages of the ZNF technology are that non-homologous end addition to suppress a gene or locus through deletions of up to a few kb. can also provide a target site for homologous integration (Cui et al., 2011, Nat Biotechnol 29:64-67). The human heavy chain antibodies produced in UniRat™ are referred to as UniAbs ^™ and can bind epitopes that cannot be attacked by conventional antibodies. Their high specificity, affinity and small size make them ideal for mono- and multi-specific applications.

In addition to UniAbs ^™ , in particular, heavy chain-only antibodies lacking camel backbone and VHH mutations and their functional VH regions are included herein. Such heavy chain-only antibodies can, for example, be produced in transgenic rats or mice that contain fully human heavy chain-only gene loci, as described, for example, in WO 2006/008548, but other transgenic mammals can also be used. such as rabbit, guinea pig, rat, rats and mice are preferred. Heavy chain-only antibodies, including their functional VHH or VH fragments, can also be produced using recombinant DNA technology by expressing the encoding nucleic acid in a suitable eukaryotic or prokaryotic host, including, for example, mammalian cells (e.g., CHO cells), intestinal stick or yeast.

Heavy chain-only antibody domains combine the advantages of antibodies and small molecule drugs: they can be mono- or multivalent; have low toxicity; and cost effective to manufacture. Because of their small size, these domains are easy to administer, including oral or topical administration, and are highly stable, including gastrointestinal stability; and their half-life can be adapted to the desired application or indication. In addition, the VH and VHH domains of HCAb can be obtained in a cost-effective manner.

In a specific embodiment, the heavy chain-only antibodies of the invention, including UniAbs™, have a native amino acid residue at the first position of the FR4 region (amino acid position 101 according to the Kabat numbering system) substituted with another amino acid residue that is capable of degrading the surface-exposed hydrophobic patch, containing or linked to a native amino acid residue at that position. Such hydrophobic patches are usually hidden at the border with the antibody light chain constant region, but become exposed to the surface in the HCAb and, at least in part, to unwanted aggregation and association of the HCAb light chain. The substituted amino acid residue is preferably charged, and more preferably positively charged, such as lysine (Lys, K), arginine (Arg, R) or histidine (His, H), preferably arginine (R). In a preferred embodiment, heavy chain-only antibodies derived from transgenic animals contain a Trp-Arg mutation at position 101. The resulting HCAbs preferably have high antigen-binding affinity and physiological solubility in the absence of aggregation.

Within the scope of the present invention, anti-CD22 heavy chain human IgG antibodies with unique sequences from UniRat™ (UniAb ^™ ) animals have been identified that bind human CD22 in a protein-cell binding ELISA (recombinant extracellular domain of CD22). The heavy chain variable region (VH) sequences identified (see FIG. 2) are positive for human CD22 protein binding and/or for binding to CD22+ cells, and all are negative for binding to cells that do not express CD22 .

The antibodies described herein bind CD22-positive Burkitt's lymphoma in the Daudi cell line (ATCC® CCL-213™), and some cross-react with the cynomolgus monkey CD22 protein. In addition, they can be designed to be cross-reactive with the CD22 protein of any animal species, if desired.

Anti-CD22 antibodies containing only heavy chains, such as UniAbs ^™ , may have an affinity for CD22 with a Kd of about 10 ^-6 to about 10 ^-11 , including, but not limited to, about 10 ^-6 to about 10 ^-10 ; from about 10 ^-6 to about 10 ^-9 ; from about 10 ^-6 to about 10 ^-8 ; from about 10 ^-8 to about 10 ^-11 ; about 10 ^-8 to about 10 ^-10 ; from about 10 ^-8 to about 10 ^-9 ; from about 10 ^-9 to about 10 ^-11 ; from about 10 ^-9 to about 10 ^-10 ; or any value in those ranges. The choice of affinity can be confirmed by a biological assessment for modulation, eg blocking, CD22 biological activity, including in vitro assays, preclinical models and clinical trials, and an assessment of potential toxicity.

Heavy chain-only antibodies that bind to non-overlapping epitopes on the CD22 protein, such as ^UniAbs.TM . known antibodies that bind to the target antigen and the antibody of interest. Using this approach, it is possible to separate a set of antibodies into those that compete with the reference antibody and those that do not. Non-competitive antibodies are identified as binding to a single epitope that does not overlap with the epitope associated with the reference antibody. Often one antibody is immobilized, the antigen binds, and a second labeled (eg, biotinylated) antibody is tested in an ELISA for the ability to bind the captured antigen. It can also be performed using Surface Plasmon Resonance (SPR) platforms including ProteOn XPR36 (BioRad, Inc), Biacore 2000 and Biacore T200 (GE Healthcare Life Sciences), and MX96 SPR tomograph (Ibis technologies BV), as well as biolayer interferometric platforms such as Octet Red384 and Octet HTX (ForteBio, Pall Inc). See the examples in this document for further details.

Typically, an antibody "competes" with a reference antibody if it causes a reduction in the binding of the reference antibody to the target antigen by about 15-100%, as determined by standard methods such as the competitive binding assays described above. In various embodiments, the relative inhibition is at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least at least about 80%, at least about 85%, at least about 90%, at least about 95% or higher.

Pharmaceutical compositions, uses and methods of treatment.

Another aspect of the present invention is the provision of pharmaceutical compositions containing one or more antibodies of the present invention in admixture with a suitable pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers used herein include, but are not limited to, adjuvants, solid carriers, water, buffers, or other carriers used in the art for storage of therapeutic components or combinations thereof.

In one embodiment, the pharmaceutical composition contains a heavy chain-only antibody (eg, UniAb ^™ ) that binds to CD22. In another embodiment, the pharmaceutical composition comprises a multispecific (including bispecific) heavy chain-only antibody (eg, UniAb™) with a binding specificity for two or more non-overlapping epitopes on the CD22 protein. In a preferred embodiment, the pharmaceutical composition comprises a multispecific (including bispecific) heavy chain-only antibody (e.g., UniAb ^™ ) with a binding specificity for CD22 and a binding specificity for a binding target on an effector cell (e.g., a binding target on a T cell, such as, for example, the CD3 protein on a T cell).

The pharmaceutical composition of antibodies used in accordance with the present invention is prepared for storage by mixing proteins having the desired purity with optional pharmaceutically acceptable carriers, excipients, or stabilizers (see, for example, Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. ( 1980)), for example, in the form of lyophilized compositions or aqueous solutions. Acceptable carriers, excipients or stabilizers are non-toxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citrate and other organic acids; 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 m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; 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 complexes (eg Zn-protein complexes); and/or non-ionic surfactants such as TWEE™, PLURONICS™ or polyethylene glycol (PEG).

Pharmaceutical compositions for parenteral administration are preferably sterile and substantially isotonic and are manufactured under the terms of the Good Manufacturing Practice (GMP). Pharmaceutical compositions may be provided in unit dosage form (eg, unit dosage form). A suitable dosage form depends on the chosen route of administration. The antibodies described herein may be administered by intravenous injection or infusion, or subcutaneously. For injection, the antibodies described herein can be formulated in aqueous solutions, preferably in physiologically compatible buffers, to reduce discomfort at the injection site. The solution may contain carriers, excipients or stabilizers as described above. Alternatively, the antibodies may be lyophilized for mixing with a suitable vehicle, such as sterile, pyrogen-free water, prior to use.

Antibody formulations are disclosed, for example, in US Pat. No. 9,034,324. Similar formulations can be used for heavy chain-only antibodies, including UniAbs ^™ , of the present invention. Antibody formulations for subcutaneous administration are described, for example, in US Pat. No. 20160355591 and US Pat.

Application methods

The anti-CD22 heavy chain-only antibodies, multispecific antibodies, and pharmaceutical compositions described herein can be used to treat diseases and conditions characterized by CD22 expression, including, without limitation, the conditions and diseases described hereinafter. Aspects of the present invention also relate to the use of the antibody described herein in the manufacture of a medicament for the treatment of a CD22-expressing B-cell disorder. Aspects of the present invention also relate to an antibody described herein for use in the treatment of a B cell disorder characterized by CD22 expression.

CD22 is a 135 kDa type I transmembrane protein that is expressed at low levels in pre- and immature B cells, maximally in mature B cells, and ultimately downregulated in plasma cells. (For example, Walker et al., Immunology, 2008 Mar; 123(3) 314-25). CD22 is highly expressed in follicular (primary and secondary B-cells), mantle and marginal B-cells and has been reported to be present in 60-80% of specimens from patients with B-cell malignancies (Alderson et al., Clin Cancer Res 2009;15(3) February 11, 2009). Because of its observed expression in a number of hematological malignancies, CD22 is a promising target for antibody-based therapies.

In one aspect, anti-CD22 heavy chain-only antibodies (e.g., UniAbs ^™ ) and pharmaceutical compositions of this invention can be used to treat hematologic malignancies characterized by CD22 expression, including, but not limited to, diffuse large B-cell lymphoma (DLBCL) , non-Hodgkin's lymphoma, B-cell chronic lymphocytic leukemia (CLL), and B-cell acute lymphoblastic leukemia (ALL).

Diffuse large B-cell lymphoma (DLBCL or DLBCL) is the most common form of non-Hodgkin's lymphoma in adults (Blood 1997 89 (11): 3909-18), with an estimated annual incidence of 7 to 8 cases per 100,000 people per year in the US and UK. It is characterized as an aggressive cancer that can occur in almost any part of the body. The causes of DLBCL are not well understood and may be caused by normal B cells as well as malignant transformation of other types of lymphoma or leukemia cells. Treatment approaches typically include chemotherapy and radiation, and as a result, overall five-year survival averages about 58% for adults. Although some monoclonal antibodies have shown promise in the treatment of DLBCL, consistent clinical efficacy has not yet been conclusively demonstrated. Therefore, there is a great need for new therapies, including immunotherapy, for the treatment of DLBCL.

In another aspect, heavy chain-only anti-CD22 antibodies (e.g., UniAbs™) and pharmaceutical compositions of this invention can be used to treat autoimmune disorders characterized by pathogenic B cells that express CD22, including, but not limited to, systemic lupus erythematosus. (SLE), rheumatoid arthritis (RA) and multiple sclerosis (MS).

Effective dosages of the compositions of this invention for treating a disease will vary depending on many different factors, including routes of administration, destination, physiological state of the patient, whether the patient is human or animal, whether other drugs are being administered, and whether the treatment is prophylactic or therapeutic. Typically, the patient is a human, but non-human mammals can also be treated, for example, pets such as dogs, cats, horses, etc., laboratory mammals such as rabbits, mice, rats, etc. , etc. Dosages can be adjusted to optimize safety and efficacy.

Dosage levels can be readily determined by one of ordinary skill in the art and can be changed as needed, for example, if the subject's response to therapy needs to be altered. The amount of active ingredient that can be combined with carrier materials to form a unit dosage form varies depending on the host being treated and the particular route of administration. Standard dosage forms usually contain from about 1 to about 500 mg of the active ingredient.

In some embodiments, the therapeutic dosage of the agent may be from about 0.0001 to 100 mg/kg, and more typically from 0.01 to 5 mg/kg, based on the body weight of the host. For example, the dosage may be 1 mg kg body weight or 10 mg/kg body weight, or in the range of 1-10 mg/kg. An exemplary treatment regimen includes administration once every two weeks, or once a month, or once every 3 to 6 months. Therapeutic substances of the present invention are usually administered several times. Intervals between single doses may be weekly, monthly or yearly. The intervals may also be irregular, as indicated by measuring the level of the therapeutic agent in the patient's blood. Alternatively, the therapeutic agents of the present invention may be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency will vary depending on the half-life of the polypeptide in the patient.

Typically, the compositions are prepared as injectable preparations, either as liquid solutions or suspensions; also can be obtained solid forms suitable for dissolution or suspension in liquid media prior to injection. The pharmaceutical compositions described herein are suitable for intravenous or subcutaneous administration, directly or after reconstitution from a solid (ie, lyophilized) composition. The drug may also be emulsified or encapsulated in liposomes or microparticles such as polylactide, polyglycolide or copolymer to enhance the adjuvant effect as described above. Langer, Science 249: 1527, 1990 and Hanes, Advanced Drug Delivery Reviews 28: 97-119, 1997. The substances of this invention may be administered in the form of a depot injection or implant formulation, which may be formulated to provide a prolonged or pulsatile release of the active substance. Pharmaceutical compositions are generally formulated as sterile, substantially isotonic, and fully compliant with all requirements of the U.S. Food and Drug Administration's GMP regulations.

The toxicity of the antibodies and antibody structures described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, by determining LD ₅₀ (dose lethal to 50% of the population) or LD ₁₀₀ (dose lethal to 100% of the population ). The dose ratio between toxic and therapeutic effect is a therapeutic indicator. The data obtained from these cell culture assays and animal studies can be used to formulate a dosage range that is not toxic for use in humans. The dosage of the antibodies described herein is preferably in the range of circulating concentrations that include an effective dose with little or no toxicity. The dosage may vary within this range depending on the dosage form used and the route of administration used. The exact composition, route of administration and dosage can be selected individually by the doctor, taking into account the condition of the patient.

Compositions for administration typically contain an antibody or other ablative agent dissolved in a pharmaceutically acceptable carrier, preferably an aqueous carrier. Various aqueous vehicles may be used, such as buffered saline and the like. These solutions are sterile and generally free of unwanted substances. These compositions may be sterilized by conventional, well known sterilization techniques. The compositions may contain pharmaceutically acceptable excipients necessary to bring them closer to physiological conditions, such as pH adjusting agents and buffering agents, toxicity adjusting agents and the like, for example sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. . The concentration of the active agent in these formulations can vary widely and will be selected primarily based on fluid volumes, viscosity, body weight, and the like, in accordance with the particular route of administration chosen and the needs of the patient (e.g., Remington's Pharmaceutical Science ( ^15th ed., 1980) and Goodman & Gillman, The Pharmacological Basis of Therapeutics (Hardman et al., eds., 1996)).

Also within the scope of the invention are kits containing active agents and formulations thereof, inventions and instructions for use. The kit may additionally contain at least one additional component, for example, a chemotherapy drug, etc. The kits usually include a label indicating the intended use of the contents of the kit. The term "label" as used in this document includes any written or recorded materials supplied with or with it, or otherwise accompanying the kit.

Now that the invention has been fully described, it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the spirit or scope of the invention.

EXAMPLES

Materials and methods

Binding of the CD22 protein

Kinetic binding experiments to determine antigen-antibody affinity were performed on an Octet QK-384 system (ForteBio) using two-layer interferometry. Anti-human IgG Fc Capture (AHC) biosensors (Forte Bio, no: lots 18-5064) were hydrated in assay buffer (1x PBS, 0.1% BSA, 0.02% Tween-20, pH 7.2) and pretreated in 100 mM glycine pH 1.5. The baseline was set in the assay buffer at 120 seconds. Biosensors AHC then immobilized with UniAbs ^TM at a concentration of 5 μg/ml for 120 seconds. Another baseline (120 seconds) was set in the assay buffer. They were then immersed in a 7-point 1:2 dilution series of human CD22 protein in assay buffer starting at 250 nM. The last well of the analyte column contained only assay buffer to check for non-specific binding between buffer and loaded biosensors and was used as a control well. Association was observed within 600 seconds, followed by dissociation within 900 seconds. Data analysis was performed using Octet Data Analysis v9.0 (ForteBio). Binding kinetics were analyzed using a 1:1 binding model.

Binding of CD22 cells

Binding to CD22 positive cells was assessed by flow cytometry (Guava easyCyte 8HT, EMD Millipore) using a Daudi cell line (ATCC). Briefly, 100,000 target cells were stained with a series of dilutions of purified UniAbs ^TM for 30 minutes at 4°C. After incubation, cells were washed twice with flow cytometry buffer (1X PBS, 1% BSA, 0.1% NaN ₃ ) and stained with goat F(ab') ₂ anti-human IgG conjugated to R-phycoerythrin (PE) (Southern Biotech, 2042-09) to detect cell-bound antibodies. After a 20 minute incubation at 4° C., the cells were washed twice with flow cytometry buffer and then the mean fluorescence intensity (MFI) was measured by flow cytometry. EC50 values were calculated using GraphPad Prism 7. Binding to CD22 positive cynomolgus monkey cells was determined using the same protocol with the following modifications: target cells were CHO cells stably transfected to express the cynomolgus CD22 extracellular domain, and each antibody was tested in one concentration (~1.7 µg/mL) so EC50 values were not calculated.

Example 1 Genetically Engineered Rats Expressing Heavy Chain Only Antibodies

The human and rat IgH locus was constructed and assembled from several parts. This involved modifying and merging the rat C-region genes downstream of the human J _H and then adding the human D-segment upstream of the V _H 6 region. Two BACs with separate human V _H gene clusters [BAC6 and BAC3] were then co-administered with a BAC named Georg encoding an assembled and modified region including human V _H 6, all D, all J _H and modified Cγ2a/1/2b ( ΔC _H 1) rats.

Were obtained transgenic rats carrying artificial heavy chain immunoglobulin loci in a disordered configuration. IgG2a(ΔC _H 1), IgG1(ΔC _H 1), IgG2b(ΔC _H 1) genes were absent in the C _H 1 segment. The IgE, IgA and 3' enhancer constant region genes were included in BAC Georg. RT-PCR and serum analysis (ELISA) of transgenic rats revealed productive rearrangement of transgenic immunoglobulin loci and expression in serum of only heavy chain antibodies of various isotypes. Transgenic rats were crossed with rats with mutated endogenous heavy chain and light chain loci previously described in US Patent Publication 2009/0098134 A1. Analysis of these animals demonstrated inactivation of rat immunoglobulin heavy and light chain expression and high expression of heavy chain-only antibodies with the variable regions encoded by the human V, D and J genes. Immunization of transgenic rats resulted in serum responses with high titers of antigen-specific antibodies containing only heavy chains. These transgenic rats expressing heavy chain-only antibodies with a human VDJ region were named UniRats™.

Example 2: Immunization

Immunization with the recombinant extracellular domain of CD22.

Twelve UniRat animals (6 HC27, 6 HC28) were immunized with recombinant human CD22 protein. Animals were immunized according to the standard protocol using Titermax/Alhydrogel adjuvant. The recombinant CD22 extracellular domain was purchased from R&D Systems and diluted with sterile saline and combined with an adjuvant. The immunogen was combined with adjuvants Titermax and Alhydrogel. The first immunization (priming) with the Titermax immunogen was performed on the left and right legs. Subsequent booster immunizations were carried out in the presence of Alhydrogel and three days before the selection of boosts were carried out with immunogen in PBS. Serum was collected from rats at the last blood draw to determine serum titers.

Serum titer results

A summary of serum titer is shown in FIG. 6. In the graphs depicted in FIG. 6, each line represents a different animal. The legend of the graphs shows the identification number of each individual animal. Binding activity for a series of 8-point dilution sera was tested by ELISA against huCD22+ Fc protein, huCD22+His-tag, CD22+His-tag rhesus monkey protein, and non-His-tag target protein. Among this group of animals, a range of levels of serum reactivity was observed for both the human CD22 protein and the rhesus monkey. A serum response to the His-tag of the protein was also observed.

Example 3 Binding to CD22 Expressing Cell Lines

On FIG. 4 shows the target binding activity of anti-CD22 heavy chain antibodies as described herein. Column 1 shows the identification number (ID) of the anti-CD22 antibody clone containing heavy chains only. Column 2 shows protein binding affinity (KD) measured in molarity. Column 3 shows the protein binding dissociation constant (K-off rate) measured in seconds. Column 4 shows Daudi cell binding measured as fold over MFI background signal. Column 5 shows binding to CHO cells stably expressing cynomolgus monkey CD22, measured as fold over baseline MFI signal. Column 6 shows binding to CHO cells that do not express CD22, measured as fold over baseline MFI signal.

Example 4 Bispecific Antibody Mediated Killing of Human Tumor Cells by Redirecting Activated T Cells

Three different CD22-positive Burkitt's lymphoma tumor cell lines (Daudi, Raji and Ramos) were stained and incubated with increasing amounts of the bispecific antibody in the presence of pre-activated human T cells. The bispecific antibody consisted of an anti-CD3 binding arm paired with an anti-CD22 VH binding domain, as shown schematically in FIG. 5D. The negative control antibody contained a VH binding domain that did not bind to CD22. CD22-negative K562 cells did not show specific lysis (data not shown). Data from three bispecific antibodies containing three different heavy chain-only anti-CD22 binding domains paired with the same anti-CD3 binding domain are shown in FIG. 5A was compared to a negative control and demonstrated antibody-mediated killing of CD22-positive Daudi tumor cells by redirecting activated T cells. Data for two bispecific antibodies containing the same heavy chain-only anti-CD22 binding domain paired with two different anti-CD3 binding domains are shown in FIG. 5B was compared to a negative control and demonstrated antibody-mediated killing of CD22-positive Raji tumor cells by redirecting activated T cells. Data for two bispecific antibodies containing the same heavy chain-only anti-CD22 binding domain paired with two different anti-CD3 binding domains are shown in FIG. 5C was compared to a negative control and demonstrated antibody-mediated killing of CD22-positive Ramos tumor cells by redirecting activated T cells.

While preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided for the purpose of illustration only. Many variations, changes and substitutions will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. It should be understood that various alternative embodiments of the present invention described herein may be used in the practice of the present invention. It is intended that the following claims define the scope of the invention and that methods and structures are thus covered within the scope of the claims and their equivalents.

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SEQUENCE LIST

<110> TENEOBIO, INC.

ALDRED, SHELLEY FORCE

<120> ANTIBODIES CONTAINING ONLY HEAVY CHAINS WHICH

BIND TO CD22

<130> TNO-0009-WO

<140> PCT/US2018/067299

<141> 21.12.2018

<150> 62/609 759

<151> 12/22/2017

<160> 87

<170> PatentIn version 3.5

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Gly Gly Ser Ile Ser Ser Gly Asp Tyr Tyr

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Gly Gly Ser Phe Ser Gly Tyr Tyr

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Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser

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Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser

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Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser

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Ala Arg Glu Asp Ser Ser Ser Trp Arg Ser

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Ala Arg Asp Asp Ser Ser Asn Trp Arg Ser

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Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

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Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

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<210> 25

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<212> Protein

<213> Artificial sequence

<220>

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Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

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Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Glu Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

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Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

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Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 26

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<212> Protein

<213> Artificial sequence

<220>

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<223> /note="Description of the artificial sequence:

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Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

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Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 27

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

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Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 28

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

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Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 29

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

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<400> 29

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 30

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

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<400> 30

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 31

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 31

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Gly Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 32

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 32

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 33

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 33

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 34

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 34

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 35

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 35

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Glu Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 36

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 36

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 37

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 37

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 38

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 38

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 39

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 39

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 40

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 40

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 41

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 41

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 42

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 42

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 43

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 43

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 44

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 44

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Glu Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 45

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 45

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 46

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 46

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 47

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 47

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Gly Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 48

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 48

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg His Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 49

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 49

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 50

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 50

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 51

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 51

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 52

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 52

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 53

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 53

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Thr Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 54

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 54

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 55

<211> 116

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 55

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr

20 25 30

Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser Leu Lys

50 55 60

Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe Ser Leu

65 70 75 80

Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Thr

85 90 95

Arg Glu Asp Ser Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr Leu Val

100 105 110

Thr Val Ser Ser

115

<210> 56

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 56

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Ala Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 57

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 57

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 58

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 58

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 59

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 59

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 60

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 60

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 61

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 61

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 62

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 62

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 63

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 63

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 64

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 64

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 65

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 65

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 66

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 66

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 67

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 67

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Thr Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 68

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 68

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 69

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 69

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 70

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 70

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 71

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 71

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 72

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 72

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Met Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 73

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 73

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 74

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 74

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Ser Pro Glu Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Lys Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 75

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 75

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 76

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 76

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Val Tyr Tyr Thr Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 77

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 77

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Phe Arg His Pro Pro Gly Lys Gly Leu Asp

35 40 45

Trp Ile Gly Ser Ile His Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 78

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 78

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser His Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 79

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 79

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 80

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 80

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 81

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 81

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ile Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Ala Tyr Tyr His Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Ile Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 82

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 82

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 83

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 83

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

20 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Ser

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 84

<211> 118

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic polypeptide"

<400> 84

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asp Asn

20 25 30

Ser His Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe

65 70 75 80

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 85

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic peptide"

<220>

<221> OPTION

<222> (2)..(2)

<223> /replace="Gly"

<220>

<221> OPTION

<222> (5)..(5)

<223> /replace="Thr" or "Ile" or "Asn"

<220>

<221> OPTION

<222>(6)..(6)

<223> /replace="Asp"

<220>

<221> OPTION

<222>(7)..(7)

<223> /replace="Ser" or "Asn"

<220>

<221> OPTION

<222>(8)..(8)

<223> /replace="Gly" or "Ser"

<220>

<221> OPTION

<222>(9)..(9)

<223> /replace="His"

<220>

<221> SITE

<222> (1)..(10)

<223> /note="Variant residues given in

sequences are not preferred

compared to those given in the footnotes for alternative

positions"

<400> 85

Gly Asp Ser Ile Ser Ser Gly Asp Tyr Tyr

1 5 10

<210> 86

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic peptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace="Val"

<220>

<221> OPTION

<222> (2)..(2)

<223> /replace="His"

<220>

<221> OPTION

<222> (4)..(4)

<223> /replace="Thr"

<220>

<221> OPTION

<222>(6)..(6)

<223> /replace="Val" or "Ser"

<220>

<221> OPTION

<222>(7)..(7)

<223> /replace="Ala"

<220>

<221> SITE

<222> (1)..(7)

<223> /note="Variant residues given in

sequences are not preferred

compared to those given in the footnotes for alternative

positions"

<400> 86

Ile Tyr Tyr Ser Gly Ala Thr

15

<210> 87

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<221> source

<223> /note="Description of the artificial sequence:

Synthetic peptide"

<220>

<221> OPTION

<222> (1)..(1)

<223> /replace="Ala" or "Lys"

<220>

<221> OPTION

<222> (3)..(3)

<223> /replace="Glu"

<220>

<221> OPTION

<222>(7)..(7)

<223> /replace="Ser"

<220>

<221> SITE

<222> (1)..(10)

<223> /note="Variant residues given in

sequences are not preferred

compared to those given in the footnotes for alternative

positions"

<400> 87

Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser

1 5 10

<---

Claims (34)

1. A heavy chain-only antibody that binds to CD22 containing a heavy chain variable region containing (a) the CDR1 sequence according to SEQ ID NO: 1, the CDR2 sequence according to SEQ ID NO: 11, and the CDR3 sequence according to SEQ ID NO: 18; or (b) the CDR1 sequence according to SEQ ID NO: 1, the CDR2 sequence according to SEQ ID NO: 12, and the CDR3 sequence according to SEQ ID NO: 19; or (c) CDR1 sequence according to SEQ ID NO: 1, CDR2 sequence according to SEQ ID NO: 12, and CDR3 sequence according to SEQ ID NO: 20. 2. The heavy chain-only antibody of claim 1, wherein the CDR sequences are contained in a human VH framework. 3. The heavy chain-only antibody of claim 1, comprising a heavy chain variable region having at least 95% sequence identity with SEQ ID NO: 24. 4. The heavy chain-only antibody of claim 3, comprising the heavy chain variable region sequence according to SEQ ID NO: 24. 5. The heavy chain-only antibody of claim 1, comprising a heavy chain variable region having at least 95% sequence identity with SEQ ID NO: 25. 6. The heavy chain-only antibody of claim 5, comprising the heavy chain variable region sequence according to SEQ ID NO: 25. 7. The heavy chain-only antibody of claim 1, comprising a heavy chain variable region having at least 95% sequence identity with SEQ ID NO: 32. 8. The heavy chain-only antibody of claim 7, comprising the heavy chain variable region sequence according to SEQ ID NO: 32. 9. The heavy chain-only antibody of claim 1, comprising a heavy chain constant region sequence in the absence of a CH1 sequence. 10. An antibody containing only heavy chains, according to any one of paragraphs. 1-9 containing the human IgG4 Fc region. 11. The heavy chain-only antibody of claim 9, wherein the heavy chain constant region comprises a CH2 domain and a CH3 domain. 12. The heavy chain-only antibody of claim 10, wherein the human IgG4 Fc region is a variant human IgG4 Fc region. 13. The heavy chain-only antibody of claim 10, wherein the human IgG4 Fc region is a silent human IgG4 Fc region. 14. CAR-T molecule that binds to CD22, containing an antibody containing only heavy chains, according to any one of paragraphs. 1–13. 15. A pharmaceutical composition containing an antibody containing only heavy chains, according to any one of paragraphs. 1-13 for the treatment of a CD22-expressing B-cell disorder, wherein the composition comprises a therapeutically effective amount of a heavy chain-only antibody. 16. A polynucleotide encoding an antibody containing only heavy chains, according to any one of paragraphs. 1–13. 17. A method of treating a B-cell disorder characterized by CD22 expression, comprising administering to a subject with said disorder an antibody containing only heavy chains, according to any one of paragraphs. 1-13 or a pharmaceutical composition according to claim 15. 18. A multispecific antibody comprising a first binding fragment that has binding specificity for CD22 and a second binding fragment that has binding affinity for an effector cell, wherein the first binding fragment contains a heavy chain variable region containing: (a) the CDR1 sequence according to SEQ ID NO: 1, the CDR2 sequence according to SEQ ID NO: 11, and the CDR3 sequence according to SEQ ID NO: 18; or (b) the CDR1 sequence according to SEQ ID NO: 1, the CDR2 sequence according to SEQ ID NO: 12, and the CDR3 sequence according to SEQ ID NO: 19; or (c) CDR1 sequence according to SEQ ID NO: 1, CDR2 sequence according to SEQ ID NO: 12, and CDR3 sequence according to SEQ ID NO: 20. 19. The multispecific antibody of claim 18, which is a bispecific antibody. 20. The multispecific antibody of claim 18, wherein the second binding fragment has a binding affinity for a T cell antigen. 21. The multispecific antibody of claim 18, wherein the second binding fragment has a binding affinity for CD3. 22. A polynucleotide encoding an antibody containing only heavy chains, according to any one of paragraphs. 1-13. 23. The use of an antibody containing only heavy chains, according to any one of paragraphs. 1-13 or a multispecific antibody according to any one of paragraphs. 18-21 for the treatment of a CD22-expressing B-cell disorder comprising administering a therapeutically effective amount of an antibody or multispecific antibody. 24. The use of claim 23 wherein the B-cell disorder is non-Hodgkin's lymphoma. 25. Use according to claim 23, wherein the B-cell disorder is B-cell chronic lymphocytic leukemia. 26. The use of claim 23 wherein the B-cell disorder is follicular lymphoma. 27. The use of claim 23 wherein the disorder is selected from diffuse large B cell lymphoma (DLBCL), systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and multiple sclerosis (MS). 28. A polynucleotide encoding a multispecific antibody according to any one of paragraphs. 18-21.

Images