TW201247221A - Improved immunotherapy - Google Patents

Abstract

The present invention provides combinations of (a) an immunoconjugate comprising at least one antigen-binding moiety and an effector moiety, and (b) an antibody engineered to have increased effector function, for use in treating a disease in an individual in need thereof. Further provided are pharmaceutical compositions comprising the combinations, and methods of using them.

Description

201247221 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates substantially to immunotherapy. More specifically, the present invention relates to immunoconjugates and engineered antibodies for use as a targeted antigen for use as an immunotherapeutic agent. Furthermore, the present invention relates to pharmaceutical compositions comprising such immunological compounds in combination with such antibodies and methods of using the same to treat diseases. [Prior Art] It is often desirable to selectively destroy individual cells or specific cell types in a variety of clinical settings. For example, the primary goal of cancer therapy is to specifically destroy tumor cells while maintaining healthy cells and tissues intact and intact. An attractive way to achieve this is to attack and destroy tumor cells by immune effector cells such as natural killer (NK) cells or cytotoxic tau lymphocytes (CTL) by inducing an immune response against the tumor. Effector cells can be activated by a variety of stimuli, including many cytokines that induce signalling events by binding to their receptors on the surface of immune cells. For example, interleukin-2 (IL 2) particularly stimulates the proliferation and activation of cytotoxic tau cells and NK cells, and has been approved for the treatment of metastatic renal cell carcinoma and malignant melanoma. However, rapid blood clearance and lack of tumor specificity require systemic administration of high doses of cytokines to achieve a sufficiently high concentration of cytokines at the tumor site to activate the immune response or have an anti-tumor effect. This high systemic content of cytokines can lead to severe toxic and noxious reactions, as is the case with IL-2. Therefore, it is necessary to specifically deliver cytokines to a tumor or tumor microenvironment when used in cancer therapy. 161561.doc 201247221 This goal can be achieved by binding cytokines to tumor antigen-specific targeting moieties such as antibodies or antibody fragments. Another advantage of such immunoconjugates is that they increase serum half-life compared to unconjugated cytokines. Its ability to maximize immunostimulatory activity at the tumor site at the lower dose while minimizing systemic side effects makes cytokine immunoconjugates the best immunotherapeutic. Another way to activate effector cells is to bind the activated Fc receptor on its surface by the Fc portion comprising the region's immunoglobulin or recombinant fusion protein. The so-called effector function of the antibody mediated by its Fc region is an important mechanism of action in antibody-based cancer immunotherapy. When an antibody that binds to the cell surface interacts with an Fc receptor on NK cells, it induces antibody-dependent cell-mediated cytotoxicity, i.e., NK cells destroy antibody-coated target cells (e.g., tumor cells). NK cells exhibit FcyRIIIa (CD16a), and FcyRIIIa recognizes IgG 1 or IgG3 subclass immunoglobulins. Other effector functions include antibody-dependent cell-mediated phagocytosis (ADCP) and complement-dependent fine sputum toxicity (CDC), which vary with the class and subclass of the antibody' because different immune cell types carry different Fc receptors. a set, the Fc receptor set distinguishes different types and subtypes of immunoglobulin heavy bond domains (eg, α, δ, γ, ε, or μ heavy chain constant domains' correspond to IgA, IgD, IgE, IgG, or IgM class antibody). Various strategies have been employed to increase the effector function of antibodies. For example, Shields et al. (J Biol Chem 9 (2), 659 Bu 6604 (2001)) show that amino acid positions 298, 333 and/or 334 (EU numbering of residues) of the Fc region are substituted for improved antibodies and Binding of Fcyllla receptor and ADCC. Other anti-resistances that exhibit amino acid modification in the Fc region and exhibit improved Fc receptor binding and effector functions are described in, for example, U.S. Patent No. 6,737,056, WO 2004/063351, and WO 2004/099249. Alternatively, the Fc receptor binding and effector function can be increased by altering the glycosylation of the antibody. The most commonly used antibody in cancer immunotherapy, the IgGl type antibody has a conserved N linkage at Asn 297 in each CH2 domain of the Fc region. Glycosylation site. Two complex biantennary oligosaccharides linked to Asn 297 are buried between the CH2 domains, forming extensive contact with the polypeptide backbone, and their presence is antibody-mediated effector function (including antibody-dependent cell-mediated cytotoxicity) ADCC)) Required (Lifely et al, Glycobiology 5, 813-822 (1995); Jefferis et al.

Immunol Rev 163, 59-76 (1998); Wright and Morrison,

Trends Biotechnol 15, 26-32 (1997)). Protein engineering studies have shown that FcyRs interact with the lower hinge region of the IgG CH2 domain (Lund et al, J Immunol 157, 4963-69 (1996)). However, FcyR binding also requires the presence of oligosaccharides in the CH2 region (Lund et al, J Immunol 157, 4963-69 (1996); Wright and Morrison, Trends Biotech 15, 26-31 (1997)), indicating both oligosaccharides and peptides. Directly contribute to the interaction site or require oligosaccharides to maintain the active CH2 polypeptide configuration. Therefore, modification of the glycosylation structure can be studied as a means of increasing the affinity of IgG1 to interact with FcYR and increasing the ADCC activity of IgG1 antibodies. Umafta et al. (Nat Biotechnol 17, 176-180 (1999) and U.S. Patent No. 6,602,684 (WO 99/54342) show β(1,4)-indolylglucosyltransferase III (GnTIII; Overexpression of a glycosyltransferase that catalyzes the formation of bisected oligosaccharides) in Chinese hamster ovary (CHO) cells significantly increases the in vitro ADCC activity of antibodies produced in such cells, 161561.doc 201247221 The contents of the literature are incorporated herein by reference in their entirety. The GnTIII in the production cell line overexpresses the production of antibodies which are concentrated by halophilic oligosaccharides, which are generally not fucosylated and are of a hybrid type. If mannosidase II (Manll) is overexpressed in the production cell line in addition to GnTin, an antibody that is condensed and not hyalinylated complex oligosaccharide is obtained (Ferrara et al., Biotechn Bioeng 93). , 851-861 (2006)). Both types of antibodies exhibit a potent increase in ADCC compared to antibodies with unmodified glycans, but only a large number of N-glycans are complex antibodies that induce significant complementarity dependence. Sexual cytotoxicity (Ferrara Etc., Bi〇techn

Bioeng 93, 851-861 (2006)). The critical factor for increasing ADCC activity appears to be the elimination of trehalose from the innermost N-acetylglucosamine residue of the oligosaccharide core', thereby improving the binding of the IgG Fc domain to FcyRIIIa (Shinkawa et al, J Biol Chem 278, 3466-3473 (2003)). Other methods of producing antibodies with reduced algae saccharification include, for example, expression in alpha (1,6)-trehalyltransferase-deficient host cells (Yamane-Ohnuki et al, Biotech Bioeng 87, 614-622 (2004) Niwa et al., J Immun〇l Methods 306, 151-160 (2006)). Although the use of free cytokines, immunoconjugates or engineered antibodies has been successful in anti-cancer immunotherapy, there is a continuing need for effective and safe novel therapeutic options in cancer therapy. SUMMARY OF THE INVENTION The inventors of the present invention have discovered that these two combinations of strategies for local immune cell activation (i.e., by cytokine immunoconjugates and engineered antibodies that increase effector function simultaneously stimulate effector cells) Improved anti-cancer 161561.doc •6- 201247221 The efficacy of immunotherapy. Accordingly, the present invention provides a combination of (a) an immunoconjugate comprising at least one antigen binding portion and an effector portion and (b) an engineered antibody that increases effector function for treating a disease in an individual in need thereof. In one embodiment, the effector moiety is a cytokine. In one embodiment, the cytokine is selected from the group consisting of IL-2, GM-CSF, IFN-[alpha], and IL-12. In a particular embodiment, the effector moiety is IL-2. In another embodiment, the effect portion is IL-12. In another specific embodiment, the IL-2 effector moiety is a mutant IL-2 effector moiety comprising at least one amino acid mutation 'in particular an amino acid substitution, as compared to the unmutated IL-2 effector moiety Amino acid mutations reduce or eliminate the affinity of the mutant IL-2 effector moiety for the alpha-subunit of the IL-2 receptor' but retain the affinity of the mutant IL-2 effector moiety for the intermediate affinity IL-2 receptor. In a specific embodiment, the mutated IL-2 effector moiety comprises i, 2 or 3 at 1, 2 or 3 positions selected from positions corresponding to residues 42, residues 45 and residues 72 of human IL-2. Amino acid substitution. In a more specific embodiment, the 'mutation" effect moiety comprises three amino acid substitutions at positions corresponding to residue 42, residue 45 and residue 72 of human IL-2. In an even more specific embodiment, the mutant IL-2 effector moiety is human IL-2 comprising an amino acid substituted F42A, Y45A and L72G. In certain embodiments, the canine 1L_2 effector moiety additionally comprises an amino acid mutation at a position corresponding to position 3 of human IL-2. This mutation abolishes the 0-glycosylation site of IL-2. In a exemplified embodiment, the mutant IL-2 effector moiety comprises the amino acid sequence SEQ ID NO: 2. Too Plus In one embodiment, the effect portion is a single chain effect portion. 161561.doc 201247221 In one embodiment, the antigen binding portion is an antibody or antibody fragment. In one embodiment, the effector moiety shares an amine terminal peptide bond or a slow terminal peptide bond with the antigen binding moiety. In one embodiment, the antigen binding portion is selected from the group consisting of a Fab molecule and a scFv molecule. In one embodiment the antigen binding moiety is a Fab molecule. In another embodiment, the antigen binding portion is divided into scFv molecules. In one embodiment, the immunoconjugate comprises a first antigen binding portion and a second antigen binding portion. In one embodiment the first antigen, the hydrazone moiety and the second antigen binding moiety are independently selected from the group consisting of a Fab molecule and a scFv molecule. In one embodiment, the first antigen binding portion and the second antigen binding portion are each a Fab molecule. In another embodiment, the first antigen binding portion and the second antigen binding portion are each a scFv molecule. In one embodiment, the effector moiety shares an amine-terminal peptide bond or a carboxy-terminal peptide bond with the first antigen-binding moiety, and the second antigen-binding moiety shares an amine-based end with the effector moiety or the first antigen-binding moiety Peptide bond or carboxy terminal peptide bond. In one embodiment, the effector moiety shares an amine-terminal peptide bond with the first antigen-binding moiety and a carboxy-terminal peptide bond with the second antigen-binding moiety. In one embodiment, the immunoconjugate consists essentially of an effector moiety joined by one or more linker sequences and a first antigen binding moiety and a second antigen binding moiety. In a particular embodiment, the immunoconjugate comprises an effector moiety (particularly a single-stranded effector moiety) and a first Fab molecule and a second Fab molecule, wherein the effector moiety is joined to the amine moiety at its amine terminal amino acid a terminal of a heavy or light bond of a Fab molecule, and wherein the effector moiety is attached to the second terminal at the carboxy terminal amino acid; the amino terminus of the heavy or light chain of the Fab molecule. I61561.doc 201247221 In certain embodiments, the antigen binding portion is directed against an antigen presented on a tumor cell or in a tumor cell environment. In a specific embodiment, the antigen binding portion is directed against an A2 domain selected from the group consisting of fibroblast activation protein (FAP), Tenscin-C (TNC A1), and Tenin C (TNC) A2) An antigen of a group of fibrin-bound protein B (EDB), carcinoembryonic antigen (CEA), and melanoma-associated chondroitin proteoglycan (MCSP). In one embodiment, the increase in effector function is selected from the group consisting of increased binding to an activated Fc receptor, increased ADCC, increased ADCP, increased CDC, increased XXX, and increased cytokine secretion. In one embodiment, the effector function is increased to increase binding to the activated Fc receptor. In a specific embodiment, the activated Fc-receptor system is selected from the group consisting of FcyRIIIa, FcyRI, and FcRylla. In one embodiment, the activated Fc receptor is FcyRIIIa. In one embodiment, this effect function is increased to increase ADCC. In one embodiment, the effector function is increased by increased binding to the activated Fc receptor and increased ADCC. In one embodiment, the anti-system is engineered by introducing one or more amino acid mutations in the Fc region. In a particular embodiment, the amino acid is mutated to an amino acid substitution. In one embodiment, the anti-system is engineered by modification of glycosylation in the Fc region. In a particular embodiment, the modification of glycosylation in the Fc region is an increase in the proportion of unfucosylated oligosaccharides in the Fc region compared to unengineered antibodies. In an even more specific embodiment, the proportion of non-fucosylated oligosaccharides in the Fc region is at least 20%, preferably at least 50%, and most preferably at least 70% of the unfucosylated oligosaccharides in the Fc region. In another specific embodiment, the glycosylation modification in the Fc region is increased in proportion to the bisected oligosaccharide in the Fc region compared to the antibody unengineered 161561.doc 201247221. In an even more specific embodiment, the proportion of quaternary oligosaccharides in the Fc region is increased by at least about 20° per aliquot of the oligosaccharide in the region. Preferably at least 5%, and most preferably at least 70%. In another specific embodiment, the glycosylation modification in the Fc region is increased in proportion to the bisected non-fucosylated oligosaccharide in the region compared to the unengineered antibody. There are at least about 25%, at least about 35%, or at least about 50% aliquot-type unfucosylated oligosaccharides in the region. In a particular embodiment, the antibody is engineered to increase the ratio of unfucosylated oligosaccharides in the Fc region compared to unengineered antibodies. The increase in the ratio of the antibody to the unfucosylated oligosaccharide in the region causes an increase in the effector function of the antibody, particularly an increase in ADCC. In a particular embodiment, the unfucosylated oligosaccharide is a bisected non-fucosylated oligosaccharide. In one embodiment, the antibody is a full length IgG class antibody, particularly an IgGl subclass antibody. In certain embodiments, the anti-system is directed against an antigen presented on a tumor cell. In a specific embodiment, the anti-system is directed to a cell selected from the group consisting of CD20, epidermal growth factor receptor (EGFR), HER2, HER3, insulin-like growth factor 1 receptor (IGF-1R), c-Met, CUB-containing domain An antigen of a group of protein 1 (CDCP1), carcinoembryonic antigen (CEA), and melanoma-associated chondroitin proteoglycan (MCSP). In a specific embodiment, the antibody is an anti-CD20 antibody engineered to increase the ratio of unfucosylated oligosaccharides in the Fc region compared to an unengineered antibody. Suitable anti-CD20 antibodies are described in WO 2005/044859, which is incorporated herein in its entirety by reference. In another specific embodiment, the antibody is engineered to be compared to an unengineered antibody. 161561.doc •10·201247221 An anti-EGFR antibody that increases the ratio of unfucosylated oligosaccharides in the Fc region. Suitable anti-EGFR antibodies are described in WO 2006/082515 and WO 2008/017963, each of which is incorporated herein by reference in its entirety. In another specific embodiment, the antibody is an anti-IGF-1R antibody engineered to increase the ratio of unfucosylated oligosaccharides in the Fc region compared to an unengineered antibody. Suitable anti-IGF-1R antibodies are described in WO 2008/077546, which is incorporated herein in its entirety by reference. In another specific embodiment, the antibody is an anti-CEA antibody engineered to increase the ratio of unfucosylated oligosaccharides in the Fc region compared to an unengineered antibody. Suitable anti-CEA antibodies are described in PCT Publication No. WO 2011/023787, the disclosure of which is incorporated herein in its entirety. In another specific embodiment, the antibody is an anti-HER3 antibody engineered to increase the ratio of unfucosylated oligosaccharides in the Fc region compared to unengineered antibodies. Suitable anti-HER3 antibodies are described in PCT Publication No. WO 2011/076683, the disclosure of which is incorporated herein in its entirety. In another specific embodiment, the antibody is an anti-CDCP1 antibody engineered to increase the ratio of unfucosylated oligosaccharides in the Fc region compared to an unengineered antibody. Suitable anti-CDCP1 antibodies are described in PCT Publication No. WO 2011/023389, the disclosure of which is incorporated herein in its entirety. In one embodiment, the anti-system is engineered by producing the antibody in a host cell that alters the activity of one or more glycosyltransferases, thereby being in the Fc region compared to an unengineered antibody. With modified glycosylation. In one embodiment, the anti-system is produced by the production of the antibody in a host cell having an increased activity of the aminotransferase III (GnTIII) of β(1,4)-indolylglucosole 161561.doc 11 201247221. Engineered to increase the proportion of unfucosylated oligosaccharides in the Fc region compared to unengineered antibodies. In a specific embodiment, the host cell additionally has increased alpha-mannosidase II (ManII) activity. In another embodiment, the anti-system is engineered to produce an antibody in a host cell having reduced alpha(ι,6)-trehalyltransferase activity, thereby interacting with an unengineered antibody The ratio of non-fucosylated oligosaccharides in the Fc region is increased. In one embodiment, the disease is a condition that can be treated by stimulating effector cell function. In one embodiment, the disease is a cell proliferative disorder. In a particular embodiment the disease is cancer. In a specific embodiment, the cancer is selected from the group consisting of lung cancer, colorectal cancer, kidney cancer, prostate cancer, breast cancer, head and neck cancer, ovarian cancer, brain cancer, lymphoma, white disease, and skin cancer. In one embodiment, the individual is a mammal. In a particular embodiment, the individual is a human. In another aspect, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier (immunoconjugate comprising at least one antigen-binding portion and an effector moiety, and (b) Antibodies engineered to increase effector function. The invention also encompasses the use of (a) an immunoconjugate comprising at least one antigen binding portion and an effector portion, and (b) an antibody engineered to increase effector function, for use in A medicament for treating a disease in an individual in need thereof. The invention further provides a method of treating a disease in an individual, the method comprising administering to the individual a therapeutically effective amount (the phantom comprises at least one antigen binding I61561.doc -12- 201247221 Partial and effector immunoconjugates and (b) Combination of antibodies engineered to increase effector function. The invention also provides a method of stimulating the function of an individual's effector cells, the method comprising administering to the individual for effective stimulation The amount of effector cell function (a) the immunoconjugate of at least one antigen-binding portion and the effector portion of the package 3 and (b) A combination of antibodies that are engineered to increase effector function. In another aspect, the invention provides a kit intended for the treatment of a disease 'the kit comprising (a) comprising at least one antigen in the same container or in a separate container An immunoconjugate that binds a portion and an effector moiety, (b) an antibody that is engineered to increase effector function, and (c) a package insert that optionally exists, the package insert containing instructions for using the combination therapy as a method of treating the disease Printed Instructions It is to be understood that the immunoconjugates and antibodies used in the pharmaceutical compositions, uses, methods and kits of the present invention may be incorporated, alone or in combination, in the above paragraphs for antibodies and immunoconjugates suitable for use in the present invention. [Embodiment] In a first aspect, the invention provides a combination of (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) an engineered antibody that increases effector function The combination is for treating a disease in an individual in need thereof. The invention further provides a method of treating a disease in an individual, The method comprises administering to the individual a therapeutically effective amount of a combination of (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) an antibody engineered to increase effector function. 161561.doc 13· 201247221 The invention also provides a method of stimulating the function of an individual effector cell, the method comprising administering to the individual an amount of (a) an immunoconjugate comprising at least one antigen-binding portion and an effector moiety, which is effective in stimulating effector cell function, and engineered A combination of antibodies that increase effector function. Unless otherwise defined hereinafter, the terms are used herein as generally used in the art. As used herein, the term "immunoconjugate" is meant to include at least one effector moiety. And a polypeptide molecule of at least one antigen binding portion. In certain embodiments, the immunoconjugate comprises at least one effector moiety and at least two antigen binding portions. A particular immunoconjugate according to the invention consists essentially of an effector moiety and two antigen-binding moieties joined by one or more linker sequences. The antigen binding portion can be joined to the effect portion by a variety of interactions and in a variety of configurations as described herein. Antigen-binding portion

161561.doc As used herein, the term "polypeptide molecule ε, γ or μ to the antigenic determinant. Suitable for light chain constant region-14-201247221 includes any of the following two isotypes: < and. As used herein, the term "control antigen binding portion" refers to an antigen binding portion in which no other antigen binding portion and effector portion are present. For example, when comparing a Fab_IL2_Fab immunoconjugate as described herein to a control antigen binding moiety, the 'control antigen binding moiety is a free Fab, wherein the Fab-IL2-Fab immunoconjugate can specifically bind to the free Fab molecule. To the same antigenic determinant. As used herein, the term "antigenic determinant" is synonymous with "antigen" and "antigenic determinant" and refers to a site on a polypeptide macromolecule that binds to an antigen binding portion to form an antigen binding portion-antigen complex (eg, , a continuous stretch of amino acid or a conformational configuration consisting of different discontinuous amino acid regions). Suitable antigenic determinants can be found, for example, on the surface of tumor cells, on the surface of virus-infected cells, on the surface of other diseased cells, in serum, and/or in extracellular matrices (ECM). "Specific binding" means selective for antigen binding and may differ from unwanted or non-specific interactions. The ability of the antigen binding moiety to bind to a specific antigenic determinant can be measured by enzyme-linked immunosorbent assay (ELISA) or other techniques well known to those skilled in the art, such as surface plasmon resonance techniques (in BIAcore instruments). Upper analysis) (Liljeblad et al., Glyco J 17, 323-329 (2000)) and traditional binding assays (Heeley, Endocr Res 28, 217-229 (2002)). The terms "anti-[antigen] antibody" and "antibody to [antigen]" mean an antibody which is capable of binding a respective antigen with sufficient affinity so that the antibody can be used as a diagnostic and/or therapeutic agent for targeting the antigen. In one embodiment, the anti-[Anti-161561.doc 15 201247221 pro] antibody binds to an unrelated protein by about 1 / ° / less than the binding of the antibody to the antigen. For example, as measured by radioimmunoassay (RIA). In certain embodiments, the antibody that binds to [antigen] has a dissociation constant (KD) of μΝ1, s1〇〇nM, $10 1^, $111]\4, 10.111]^, 50.0111]^ or $0.001111^ (eg 1 〇·8 Μ or less than ΙΟ·8 Μ, for example ΙΟ·8 Μ to 1〇·13 Μ, for example ΙΟ·9 Μ to 1〇·13 Μ). It should be understood that the above definition also applies to the antigen binding portion that binds to an antigen. As used herein, the terms "first" and "second" are used in relation to the antigen-binding portion and the like to facilitate differentiation in the presence of more than one type of portion. The use of such terms is not intended to impart a particular order or orientation to the immunoconjugate unless expressly stated otherwise. As used herein, the term "effector moiety" refers to a polypeptide, such as a protein or glycoprotein, that affects cellular activity (e.g., via signal transduction or other cellular pathways). Thus, the effector portion of the invention can be associated with receptor-mediated signaling. This signaling transmits signals from outside the cell membrane to modulate the response of cells with one or more receptors of the effector moiety. In one embodiment, the effector moiety elicits a cytotoxic response to a cell bearing one or more of the effector moieties. In another embodiment, the effector moiety can elicit a proliferative response of cells with one or more receptors of the effector moiety. In another embodiment, the effector moiety can initiate the differentiation of cells with receptors of the effector moiety. In another embodiment, the effector moiety can alter the expression (i.e., up- or down-regulation) of endogenous cellular proteins in cells bearing the receptor of the effector moiety. Non-limiting examples of effector moieties include cytokines, growth factors, hormones, enzymes, receptors, and cofactors. The effector moiety can be associated with the antigen-binding portion of the 161561.doc • 16 - 201247221 to form an immunoconjugate. As used herein, the term "cytokine" refers to a molecule that mediates and/or regulates a biological or cellular function or process (e.g., immunity, inflammation, and blood cell production). As used herein, the term "cytokine" includes "lymphokines", "chemokines", "monokines" and "interleukins". Examples of suitable cytokines include, but are not limited to, GM-CSF, IL-la, IL-ip, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7 > IL- 8 ' IL-10 ' IL-12 ' IFN-a > IFN-β ' IFN-γ ' MIP-la, ΜΙΡ-1β, TGF-β, TNF-a and TNF-β. Specific cytokines are IL-2 and IL-12. The term "cytokine" as used herein is also intended to include cytokine analogs comprising one or more amino acid mutations in the amino acid sequence of the corresponding wild-type cytokine, such as the IL-2 analogs described below. : Sauv6 et al, Proc Natl Acad Sci USA 88, 4636-40 (1991); Hu et al, Blood 101, 4853-4861 (2003) and U.S. Patent Publication No. 2003/0124678; Shanafelt et al, Nature Biotechnol 18 , 1197-1202 (2000); Heaton et al, Cancer Res 53, 2597-602 (1993) and U.S. Patent No. 5,229,109; U.S. Patent Publication No. 2007/0036752; WO 2008/0034473; WO 2009/061853 ; or context. As used herein, the term "single-chain" refers to a molecule in which the amino acid monomers contained are linearly linked by peptide bonds. In one embodiment, the effector moiety is a single-stranded effect moiety. Non-limiting examples of single chain effect moieties include cytokines, growth factors, hormones, enzymes, receptors, and cofactors. When the effector moiety is a cytokine and the cytokine of interest is normally present in the form of a multimeric form in nature 161561.doc 17 201247221, the individual units of the polycytokine are sequentially encoded by a single strand of the effector moiety. Thus, non-limiting examples of suitable single-stranded effect moieties include GM-CSF, IL-la, IL-lp, IL-2'IL-3, IL-4, IL-5, IL-6, IL-7, IL_8 , IL-1, IL_12, IFN_a, IFN_p, ΙρΝ γ, MIP-la, ΜΙΡ-Ιβ, TGF-β, TNF-a and TNF-β. As used herein, the term "control effect portion" refers to an unbound effect portion. For example, when comparing the Jiang 2 immunoconjugate to the control effect portion as described herein, the control effect portion is free unbound ^ 2 . Similarly, for example, when comparing the IL_12 immunoconjugate to the control effect, the control effect portion is free unbound IL-12 (e.g., in the form of a heterodimeric protein in which the P40 and P35 subunits share only the disulfide bond). The term "effector moiety receptor" as used herein refers to a polypeptide molecule capable of specifically binding to an effector moiety. For example, when IL_2 is part of the effector pool, the 'receptor-2 molecule (e.g., the immunoconjugate comprising IL_2) should be part of the receptor. Similarly, for example, the effect of the immune response is the part of the effector moiety is the receptor. When the effect: the specific binding to more than one receptor, the specific binding to the effector is that all of the receptors are "effector partial receptors" of the effector moiety. (d) "Antibody" is used broadly herein and encompasses various antibody structures including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies: such as bispecific antibodies, and antibody fragments, as long as they exhibit the desired antigen Knot. It is active and contains (iv) or an area equivalent to the immunoglobulin region. "Full length antibody", "intact antibody" and "entire antibody" are used interchangeably herein to refer to a structure 161561 having substantially similar structure to a native antibody, d〇c • 18· 201247221 or having an Fc as defined herein. The antibody of the heavy chain of the region. "Native antibody" refers to a naturally occurring immunoglobulin molecule having a different structure. For example, a native IgG antibody is a heterotetrameric protein of about 15 〇〇〇 Dalton, composed of two identical light bonds and two identical heavy bonds bonded by a disulfide bond. Each heavy bond from the N-terminus to the c-terminus has a variable region (VH), also known as a variable heavy or heavy chain variable domain, followed by three constant domains (CH1, CH2, and CH3), also known as heavy chains. Constant zone. Similarly, each light bond from the n-terminus to the c-terminus has a variable region (VL) 'also known as a variable light or light chain variable domain, followed by a constant light (CL) domain, also known as a light chain constant Area. The antibody light chain can be classified into one of two types, referred to as K and λ, based on the amino acid sequence of its constant domain. "Antibody fragment" refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds to an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab,)2, a bifunctional flb antibody, a linear antibody, an early bond antibody molecule (eg, SCFV), a single domain antibody, and A multispecific antibody formed from an antibody fragment. For a review of certain antibody fragments, see Hudson et al, Nat. Med 9, 129-134 (2003). For a review of scFv fragments, see, for example, piiickthun, Pharmacology of Monoclonal Antibodies, vol. 113,

Rosenburg and Moore, ed., Springer-Verlag, New York, pp. 269-315 (1994); see also w〇 93/16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. A discussion of Fab and F(ab')2 fragments comprising a rescue receptor binding to an antigenic determinant residue and having an increased in vivo half-life is described in U.S. Patent No. 5,869,046. A bifunctional antibody is an antibody fragment having two antigen binding sites and having a bivalent or bispecificity with 161561.doc -19· 201247221. See, for example, EP 404,097; WO 1993/01161; Hudson et al, Nat Med 9, 129-134 (2003); and Hollinger et al, Proc Natl Acad Sci USA 90, 6444-6448 (1993). Trifunctional and tetrafunctional antibodies are also described in Hudson et al, Nat Med 9, 129-134 (2003). A single domain antibody is an antibody fragment comprising all or a portion of a heavy chain variable domain or all or a portion of a light chain variable domain of an antibody. In certain embodiments, the single domain antibody is a human single domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No. 6,248,516 B1). Antibody fragments can be made by a variety of techniques including, but not limited to, proteolytic digestion of intact antibodies as described herein and production by recombinant host cells (e.g., E. coli or phage). The term "antigen binding domain" refers to a portion of an antibody that comprises a region that specifically binds to an antigen and is complementary to some or all of the antigen. The antigen binding domain can be provided, for example, by one or more antibody variable domains (also known as antibody variable regions). Specifically, the antigen binding domain comprises an antibody light chain variable region (VL) and an antibody heavy chain variable region (VH). The term "variable region" or "variable domain" refers to a domain in which an antibody binds to an antigen in a heavy or light chain of an antibody. The heavy chain variable domain and the light chain variable domain (VH and VL, respectively) of the native antibody generally have a similar structure, wherein each domain comprises four conserved framework regions (FR) and three hypervariable regions (HVR). See, for example, Kindt et al, Kuby Immunology, 6th ed., W.H. Freeman and Co., p. 91 (2007). A single VH or VL domain may be sufficient to confer antigen binding specificity. As used herein, the term "hypervariable region" or "HVR" refers to each of the antibodies that are highly variable in sequence and/or form a structurally defined loop ("hypervariable loop") in the 161S61.doc -20- 201247221 variant. region. In general, native 4-chain antibodies contain six HVRs; three in VH (HI, H2, H3) and three in VL (LI, L2, L3). HVRs generally comprise amino acid residues from the hypervariable loop and/or complementarity determining regions (CDRs), the complementarity determining regions having the highest sequence variability and/or involving antigen recognition. In addition to CDR1 in VH, the CDRs generally comprise an amino acid residue that forms a hypervariable loop. Hypervariable regions (HVRs) are also referred to as "complementarity determining regions" (CDRs), and are used interchangeably herein to refer to the variable portion of the antigen binding region. This particular region has been described by Kabat et al, US Dept, of Health and Human Services, Sequences of Proteins of Immunological Interest (1983) and Chothia et al, J Mol Biol 196: 901-917 (1987), where When compared to each other, the definitions include overlaps or subsets of amino acid residues. "However, applications involving the definition of CDRs of antibodies or variants thereof are intended to be within the scope of the terms as defined and used herein. Suitable amino acid residues encompassing the CDRs as defined in each of the references cited above are set forth in the Table 1 below for comparison. The exact number of residues that encompass a particular CDR will vary depending on the sequence and size of the CDR. Where the variable region amino acid sequence of an antibody is known, those skilled in the art will generally be able to determine which residues constitute a particular CDR. Table 1. CDR definitions 1 CDR Kabat Chothia AbM2 VH CDR1 31-35 26-32 26-35 VhCDR2 50-65 52-58 50-58 Vh CDR3 95-102 95-102 95-102 I61561.doc 21 201247221 VL CDR1 24 -34 26-32 24-34 VL CDR2 50-56 50-52 50-56 Vi. CDR3 89-97 91-96 89-97 1 Table 1 _ All CDR definitions are numbered according to Kabat et al. Rules (see below). 2 "AbM" with the lowercase letter r b" as used in Table 1 means the CDR ° as defined by Oxford Molecular's "AbM" antibody modeling software.

Kabat et al. also define a variable region sequence numbering system suitable for any antibody. Those skilled in the art will be able to explicitly assign this "Kabat numbering" system to any variable region sequence without relying on any experimental data other than the sequence itself. As used herein, "Kabat number" means

The numbering system described by Kabat, U.S. Dept, of Health and Human Services, "Sequence of Proteins of Immunological Interest" (1983). Unless otherwise specified, the reference to the position of a particular amino acid residue in the variable region of the antibody is based on the Kabat numbering system. The polypeptide sequences of the Sequence Listing (i.e., SEQ ID Nos. 3, 4, 5, 6, 7, 8, 9, etc.) are not numbered according to the Kabat numbering system. However, converting the sequence number of the sequence listing to Kabat numbering is well within the skill of the average skilled person. "Framework" or "FR" refers to a variable domain residue other than a hypervariable region (HVR) residue. The variable domain FR is generally composed of four FR domains: FR1, FR2, FR3, and FR4. Therefore, the 'HVR and FR sequences are generally present in VH (or VL) in the following order: FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4. The antibody "category" refers to the type of 161561.doc •22· 201247221 of the constant or constant region of the & There are five major antibody classes: IgA, IgD, IgE, IgG, and IgM' and several of these classes can be further divided into subclasses (isotypes), such as IgGi, IgG2, lgG3, IgG4, and IgA2. The heavy-chain constant domains corresponding to different immunoglobulin classes are called α, δ, ε, γ, and μ, respectively. The term "Fc region" is used herein to define a C-terminal region of an immunoglobulin heavy bond containing at least a portion of the constant region. The term encompasses both native sequence and variant Fc regions. Although the IgG heavy chain may vary slightly by the boundaries of the region, the human IgG heavy chain Fc region is generally defined as extending from Cys226 or from ΡΓ〇230 to the carboxy terminus of the heavy chain. However, the C-terminus of the Fc region may or may not be present in the amino acid (Lys447). Unless otherwise specified herein, the amino acid residue numbering in the Fc region or constant region is based on the EU numbering system, also known as the EU index, as in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service. , described in National Institutes of Health, Bethesda, MD, 1991. A region equivalent to an immunoglobulin Fc region" is intended to include a dual gene variant of a naturally occurring immunoglobulin Fc region and to have a function, such as an antibody, that produces a substitution, addition or deletion without substantially reducing the immunoglobulin-mediated effect. Variants of a change in the ability of a cell-mediated cytotoxicity), for example, can be deleted from the N-terminus or C-terminus of the immunoglobulin Fc region without substantial loss of biological function. Amino acid. Such variants can be selected according to the general rules known in the art to have minimal effect on activity (see, for example, B〇vvie et al., Science 247, 1306-1310 (1990)). The term "effector" is used. When used with antibodies, it can be attributed to antibodies 161561.doc •23- 201247221

The biological activity of the Fc region, which varies with the antibody isotype. Examples of antibody effector functions include: C1 q binding and complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); antibody-dependent cellular phagocytosis (ADCP); cytokines Secretion; immune complex-mediated antigens exhibit uptake of antigen by cells; downregulation of cell surface receptors (eg, B cell receptors); and activation of B cells. The term "effector cell" as used herein refers to the presence of an effector partial receptor (eg, a cytokine receptor) and/or an Fc receptor on the surface of a binding effector moiety (eg, a cytokine) and/or on the Fc region of an antibody. And help to destroy the lymphocyte population of the stem cells (such as tumor cells). Effector cells can mediate, for example, cytotoxic or purulocyte effects. Effector cells include, but are not limited to, effector T cells such as CD8+ cytotoxic T cells 'CD4+ helper T cells, γδΤ cells, sputum cells, lymphokine activation killer (LAK) cells, and giant sputum cells/monocytes. Depending on the mode of expression of the receptor, there may be a different subset of effector cells, ie (a) a receptor that expresses a particular effector moiety but not a pc receptor and is stimulated by an immunoconjugate of the invention rather than an antibody. (eg, a T cell that expresses an IL-2 receptor); (b) a cell that exhibits an Fc receptor but does not exhibit a specific effector moiety and is stimulated by an antibody of the invention, but not an immunoconjugate; and (c) expresses an Fc A receptor and a receptor of a specific effector moiety and simultaneously stimulated by the antibody of the present invention and an immunoconjugate (for example, NK cells expressing an Fcylll receptor and an IL-2 receptor). As used herein, the term "engineering" is taken to include any manipulation of the peptide backbone of a naturally occurring or recombinant polypeptide or fragment thereof or post-translational manipulation. Engineering modifications include modifying amino acid sequences, modifying glycosylation patterns, or modifying 161561.doc • 24· 201247221 The side chain groups of the amino acid, and combinations of such methods. "Engineering" (especially with the prefix "glyce (glyC0_)") and the term "glycosylation engineering" include metabolic engineering of cellular glycosylation mechanisms, including genetic manipulation of the oligosaccharide synthesis pathway to alter cells Glycosylation of glycoproteins expressed. In addition, glycosylation engineering involves the effects of mutations and cellular environment on glycosylation. In one embodiment 'glycosylation is engineered to alter glycosyltransferase activity. In a particular embodiment, engineering results in altered glucosyltransferase activity and/or trehalyltransferase activity. Glycosylation engineering can be used to obtain "host cells with increased GnTIII activity" (eg, one or more of the ones that have been manipulated to exhibit an increase in the activity of βΟ,Μ-Ν-acetylglycolyltransferase III (GnTIII) a host cell of a polypeptide), a host cell with increased r Manll activity (eg, a host cell that has been manipulated to exhibit an increased amount of one or more polypeptides having alpha-mannosidase n (Manll) activity) or "having a reduced alpha (" 1,6) Host cells for trehalyltransferase activity (e.g., host cells that have been manipulated to exhibit reduced amounts of alpha (丨, 6) trehalyl transferase). As used herein, the term "amino acid mutation" is intended to encompass amino acid substitutions, deletions, insertions, and modifications. Any combination of substitutions, deletions, insertions, and modifications can be made to obtain the final construct, with the proviso that the final construct possesses desirable characteristics, such as reduced binding to Fc receptors. Amino acid sequence deletions and insertions include deletion and insertion of amino acid at the amino terminus and/or carboxy terminus. The specific amino acid is mutated to an amino acid substitution. In order to alter, for example, the binding characteristics of the Fc region, a non-conservative amino acid substitution is particularly preferred, i.e., an amino acid is replaced with another amino acid having a different structure and/or chemical nature. Amine 161561.doc -25- 201247221 base acid substitutions include amino acid substitutions by non-naturally occurring amino acids or from naturally occurring 20 standard amino acids (eg 4-hydroxyproline, 3-A) Substituted by histidine, ornithine, homoserine, 5-hydroxyl-amino acid. Amino acid mutations can be produced using genetic or chemical methods well known in the art. Genetic methods can include site-directed mutagenesis, PCR, gene synthesis, and the like. It is contemplated that methods for altering the amino acid side chain groups, such as chemical modifications, by methods other than genetic engineering can also be used. "Percent amino acid sequence identity (° 〆 0)" relative to a reference polypeptide sequence is defined as after aligning the reference polypeptide sequence with the candidate sequence and, if necessary, introducing a gap to achieve a maximum sequence identity percentage, and without any Where a conservative substitution is considered to be part of sequence identity, the percentage of amino acid residues in the candidate sequence that are identical to the amino acid residues in the reference polypeptide sequence. The amino acid sequence identity percentage can be determined in a variety of ways within the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine parameters suitable for the alignment sequence, including any algorithms required to achieve maximum alignment over the entire length of the sequence being compared. However, for the purposes of this article 'Use the sequence comparison computer program ALIGN-2 to generate amino acid sequence identity percentage values ^ ALIGN-2 sequence comparison computer program was created by Genentech, Inc. and the source code is already with the instruction manual Archived in the US Copyright Office (Washington DC, 20559), where it is registered under the US copyright registration number τχυ51〇〇87. The ALIGN-2 program is publicly available from Genentech, Inc. (South San Francisco, California) or may be compiled from source code. ALIGN-2 161561.doc • 26 · 201247221 The formula should be compiled for UNIX operating systems, including digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and are unchanged. In the case of amino acid sequence comparison using ALIGN-2, the percent identity of the amino acid sequence of a given amino acid sequence A, with or relative to a given amino acid sequence b is calculated as follows (or may be referred to as established The amino acid sequence A has, or corresponds to, a given amino acid sequence B having or comprising a certain percentage of amino acid sequence identity):

10〇χ Fraction X/Y where X is the number of amino acid residues that are uniformly matched by the sequence alignment program aligN-2 in the program alignment of A and B, and wherein Y is the amino acid residue in B total. It will be appreciated that if the length of the amino acid sequence A is not equal to the length of the amino acid sequence B, then the percent identity of the amino acid sequence of A and B will not be equal to the percent identity of the amino acid sequence of B and A. All amino acid sequence identity percent values used herein are obtained using the ALIGN-2 computer program as described in the previous paragraph, unless otherwise expressly stated. The terms "host cell", "host cell strain" and "host cell culture" are used interchangeably and refer to cells into which an exogenous nucleic acid has been introduced, including progeny of such cells. Host cells include "transformants" and "transformed cells". Transformed cells include primary transformed cells and their progeny, regardless of the number of generations. The nucleic acid inclusions of the progeny are not identical to the parent cells, but may contain mutations. Included herein are mutant progeny that have the same function or biological activity as screened or selected in the original transformed cell. Host cells are any type of cell system that can be used to produce antibodies and immunoconjugates for use in the present invention. In one embodiment, the host cell is engineered to produce an antibody having an oligosaccharide modified by 161561.doc s -27-201247221. In certain embodiments, the host cell has been manipulated to exhibit an increased amount of one or more polypeptides having beta (丨,4)_N_ethylglucanisyltransferase III (GnTIII) activity. In certain embodiments, the host cell has been further engineered to exhibit an increased amount of one or more polypeptides having Mannosidase II (Manll) activity. Host cells include cultured cells, such as mammalian cultured cells, such as CHO cells, BHK cells, NSO cells, SP2/0 cells 'Υ〇 myeloma cells, ρ3χ63 mouse myeloma cells, PER cells, PER.C6 cells, or fusion tumors. Cells, yeast cells, insect cells, and plant cells, to name a few; and cells contained in a transgenic animal, a transgenic plant, or a cultivated plant or animal tissue. The term "polypeptide having GnTIII activity" as used herein refers to a β-mannose core capable of catalyzing the mannose-based core of an oligosaccharide in which a N-acetylglucosamine (GlcNAc) residue is linked to N by a β-, 4 linkage. A polypeptide linked to a mannoside addition. This term includes the display of β(1,4)-indolyl-glucosyltransferase 亦 (also known as β-1,4-mannosyl-glycoprotein 4-β-Ν-acetamido Glycosyl-transferase (EC 2.4.1.144), according to the Nomenclature Committee of the International

Union of Biochemistry and Molecular Bi〇l〇gy) (NC-IUBMB)) enzyme activity that is similar but not necessarily identical (eg, measured in a specific biological assay in the presence or absence of a dose-dependent) Fusion polypeptide. In the presence of a dose-dependent, it does not need to be identical to GnTIII, but is substantially similar in dose dependence to the established activity when compared to GnTIII (ie, the candidate polypeptide will exhibit greater activity or no more than GnTIII It is about 25 times or less, and preferably does not exceed about 1 〇 times the activity of 161561.doc -28-201247221, and preferably not more than about 3 times the activity). In certain embodiments, the polymorphism with GnTm activity is a fusion polypeptide comprising a catalytic domain of GnTm and a Golgi localization domain of a heterologous Golgi resident polypeptide. The localization domain is especially the localization domain of 甘__ or (4), most particularly the localization domain of mannose lipase II. Alternatively, the Golgi localization domain is selected from the group consisting of a localization domain of mannose, a localization domain of GnTII, and a localization domain of an alpha 16 core trehalyl transferase. Methods for producing such fusion polypeptides and using the same to produce an antibody having an increased effector function are disclosed in U.S. Patent Application Serial No. 60/495,142, and U.S. Patent Application Publication No. The contents of such documents are expressly incorporated herein by reference in its entirety in the entirety of the entire disclosure. The term "Golgi localization domain" as used herein refers to an amino acid sequence in the G〇igi resident polypeptide that is responsible for immobilizing the polypeptide in the Golgi c〇mplex. Typically, the localization domain comprises the amino terminus of the enzyme "tail j. The term "polypeptide having Manll activity" as used herein refers to a terminal 1,3- which is capable of catalyzing the GlcNAcMansGlcNAc2 mannose intermediate of the N-linked oligos. A polypeptide in which 1,6-linked α-D-mannose residues are hydrolyzed. This term includes the expression of Golgi a-mannosidase 11 (also known as mannosyl oligosaccharide l, 3-l,6-a-mannosidase II (EC 3.2.1.114), according to International Biochemistry and Molecular Biology A polypeptide of the Association Nomenclature Commission (NCMUBMB) that has similar activity but not necessarily the same enzymatic activity. An "activated Fc receptor" is an Fc receptor that causes a signaling event that stimulates a cell carrying a receptor to perform an effector function after ligation of the Fc region of the antibody. Activation by 161561.doc -29-201247221 including FcYRIIIa (CD16a), FcyRI (CD64), FcyRIIa (CD32) and FcaRI (CD89) 〇 antibody-dependent cell-mediated cytotoxicity (ADCC) for immune effector cell lysis The immune mechanism of antibody-coated target cells. Target cells are cells that specifically bind to an antibody or fragment thereof comprising an Fc region (generally via a protein portion that is the N-terminus of the region). The term "ADCC increase" as used herein is defined as the increase in the number of target cells that are dissolved in the surrounding medium of the target cell at a given antibody concentration by a defined ADCd at a given time and/or The concentration of antibody in the surrounding medium of the target cells required to dissolve a predetermined number of target cells by the ADCC mechanism is reduced for a given period of time. The ADCC increase is relative to ADCC mediated by the same standard production, purification, formulation, and storage methods (known to those skilled in the art) that are produced by the same type of host cell but not yet engineered by the same antibody. For example, ADCC mediated by an antibody produced by a host cell having an altered glycosylation pattern (eg, a glycosyltransferase GnTIII or other glycosyltransferase) engineered by the methods described herein. The increase is relative to ADCC mediated by the same antibody produced by the same type of unengineered host cell. "Antibody dependent cell mediated cytotoxicity (ADCC) increased antibody" means an antibody that is increased by ADCC, as determined by any suitable method known to those skilled in the art. One has accepted the in vitro ADCC analysis as follows: 1) The assay uses a marker cell known to represent the target antigen recognized by the antigen binding region of the antibody; 161561.doc • 30· 201247221 2) The analysis uses self-randomized selection of health Human peripheral blood mononuclear cells (PBMC) isolated from donor blood were used as effector cells; 3) The assay was performed according to the following protocol: i) PBMCs were isolated using standard density centrifugation procedures and suspended in RPMI at 5 X 106 cells/ml. In the cell culture medium; ii) the target cells are grown by standard tissue culture methods, having a temperature above 90°/. The index of viability was collected in the growth phase, washed with RPMI cell culture medium, labeled with 100 microcuries 51 Cr, washed twice with cell culture medium, and resuspended in cell culture medium at a density of 1 〇 5 cells/ml; Iii) Transfer 100 μl of the above final target cell suspension to each well of a 96-well microtiter plate; iv) serially dilute the antibody from 4〇〇〇ng/ml to 〇.〇4 ng/ in cell culture medium Ml, and 50 microliters of the resulting antibody solution was added to the target cells in a %-well microtiter plate, and the test was repeated three times to cover various antibody concentrations in all of the above concentration ranges; v) for the maximum release (MR) control, containing Receiving 50 μl of a 2% (V/V) aqueous solution of nonionic detergent (Nonidet, Sigma, St. Louis) instead of the antibody solution (above point) in 3 other wells in the tray of labeled target cells; W) For a spontaneous release (SR) control, 3 other wells in a dish containing labeled target cells receive 5 μl of microliters of RPMI cell culture medium instead of antibody solution (point iv above); vii) 96-well microtiter plated at 5〇xg Heart for 1 minute, and incubate for 1 hour at 4 C; 16I561.doc •31 · 201247221 νηι) Add 50 μl of PBMC suspension (point i above) to each well to produce a 25:1 effector cell: Target cell ratio, and at 5% c〇2 atmosphere / 37. Each plate was placed in an incubator for 4 hours; ijO collected cell-free supernatant from each well and quantified experimental release of radioactivity (ER) using a gamma counter; X) for each antibody concentration 'according to formula (ER-MR)/ (MR-SR) xl〇〇 Calculate the percentage of specific lysis 'where ER is the average radioactivity quantified for antibody concentration (see point ix above) and MR is quantified for the MR control group (see point v above) Average radioactivity (see point 以上 above) and is the average radioactivity quantified for the SR control group (see point Vi above) (see point ix above); 4) "ADCC increase" is defined as tested above a decrease in the maximum percentage of specific lysis observed over the range of antibody concentrations, and/or a decrease in the concentration of antibody required to achieve a maximum percentage of the specific lysis half observed over the range of antibody concentrations of the above formula . The ADCC increase is based on the same standard known to those skilled in the art to produce, purify, formulate, and store ADCCs mediated by the same type of host cell but not engineered by the same antibody. Measure. "Combination" (and grammatical variations thereof) as used herein encompasses a combination of an immunoconjugate according to the invention and an antibody, wherein the immunoconjugate and the anti-system are in the same or different containers, in the same or different pharmaceuticals Administration in a formulation, together or separately, simultaneously or sequentially in any order, and by the same or different routes, the restriction is that the immunoconjugate and the antibody can simultaneously exert their biological effects in vivo, ie At the same time, thorn 161S61.doc •32· 201247221 stimulated effector cells. For example, 'combination' of an immunoconjugate and antibody according to the invention may mean first administering an immunoconjugate in a particular pharmaceutical formulation' followed by administration of the antibody in another pharmaceutical formulation, or first administering The antibody in a particular pharmaceutical formulation is then administered to an immunoconjugate in another pharmaceutical formulation. An "effective amount" of a medicament is the amount necessary to cause a physiological change in the cell or tissue to which it is administered. A "therapeutically effective amount" of a medicament (e.g., a pharmaceutical composition) refers to an amount effective to achieve a desired therapeutic result or a preventative result after a necessary period of time. For example, a therapeutically effective amount of an agent that eliminates, reduces, delays, minimizes, or prevents an adverse effect of a disease. A therapeutically effective amount of a combination of several active ingredients can be a therapeutically effective amount of each active ingredient or, in order to reduce treatment The combination of several active ingredients in a therapeutically effective amount may be effective to produce an additive effect or an additive effect or a synergistic effect and, when combined, therapeutically effective but when used alone is a low or a few active ingredients. The amount of the individual active ingredient of the subtherapeutic am〇unt. An individual or a subject is a mammal. Mammals include, but are not limited to, domestic animals (eg, cows, sheep, cats, dogs, and horses), primates (eg, humans and non-human primates, such as monkeys), rabbits, and rodents (eg, small Rat and rat). In particular, the individual is a human being. The term "pharmaceutical composition" means a biologically active right wire that is present on the melon.

161561.doc means the active ingredient contained in it - the individual to whom the formulation will be administered -33 - 201247221

"Medicalally acceptable A &A; m & system" means that in addition to the active ingredient in a pharmaceutical formulation: = it is not toxic to the individual. A pharmaceutically acceptable carrier package a) a buffer, excipient, stabilizer or preservative. The treatment of change (and its grammatical variants) refers to a clinical intervention that attempts to alter the natural course of the disease in the individual being treated and can be performed for prophylactic purposes or in the course of clinical pathology. Ideal therapeutic effects include, but are not limited to, preventing the onset or recurrence of the disease, alleviating the symptoms, and attenuating any direct or indirect pathological consequences of the disease 'preventing metastasis, reducing the rate of disease progression, improving or ameliorating the condition of the disease, and reducing or improving the prognosis. In some embodiments, the combination of the invention is used to delay disease progression or slow disease progression. The term "package (four) page" is used to refer to the instructions commonly included in commercial packaging for therapeutic products, which contain indications relating to therapeutic products such as (4), doses, administration, combination therapy, contraindications and / or warning information. Immunoconjugates An immunoconjugate suitable for use in the present invention is a polypeptide molecule comprising at least one effector moiety and at least one antigen binding moiety. An immunoconjugate can be prepared by chemically binding an effector moiety to an antigen binding moiety, or by expressing an effector moiety and an antigen binding moiety as a fusion protein (see, for example, Nakamura and Kubo, Cancer 80, 2650-2655 (1997), and Becker is a person, 1 "〇.]^11人〇3(18(^178八93 7826-7831 (1996)). In the present invention, an immunoconjugate which is expressed as a fusion protein is generally preferred. In certain embodiments, the effector moiety shares an amine-terminal peptide bond or a carboxy-terminal peptide bond with the antigen-binding moiety (ie, the immunization 161561.doc-34.201247221 conjugate is a fusion protein). In such immunoconjugates, The effector moiety can, for example, be fused to an immunoglobulin heavy or light chain. Particularly suitable for use in the present invention is an immunoconjugate comprising an antibody fragment such as a Fab or scFv molecule as an antigen binding moiety. Exemplary antibody fragment/cytokine immunological binding The substance is described, for example, in Savage et al, Br J Cancer 67, 304_310 (1993); Yang et al, Mol. Immunol 32, 873-881 (1995); PCT Publication WO 2001/062298 A2; PCT Publication No. 5,650,150; PCT Publication No. WO 2006/119897 A2; and PCT Publication WO 99/29732 A2 ° In one embodiment, the effect portion is a single-chain effect portion. In one embodiment, the effect portion is a cytokine In one embodiment, the immunoconjugate comprises at least two antigen binding portions. The antigen binding portion and effector portion of the immunoconjugate comprise the above and below detailed description. The antigen binding portion of the immunoconjugate can be directed to a plurality of targets. Molecules (eg, antigenic determinants on protein molecules expressed on tumor cells or tumor stromas). Non-limiting examples of antigen binding moieties are described herein. Immunoconjugates as described herein are generally useful for exhibiting one or more of the following properties : high potency specificity, reduced toxicity and/or improved stability, especially when compared to immunoconjugates that target the same antigenic determinant and carry different configurations of the same effector moiety. The conjugate is further described in PCT Publication No. WO 20 11/020783, the disclosure of which is incorporated by reference in its entirety. The immunoconjugate package 161561.doc-35-201247221 described in the PCT Publication No. WO 2011/020783 contains at least two antigen binding domains. Thus, in one embodiment The intermediate immunoconjugate comprises at least a first effector moiety and at least a first antigen binding moiety and a second antigen binding moiety. In one embodiment, the first effect portion is a single bond effect portion. In one embodiment, the first antigen binding portion and the second antigen binding guanidine line are independently selected from the group consisting of SCFV molecules and Fab molecules. In a particular embodiment, the first antigen binding portion and the second antigen binding portion are each a Fab molecule. In another embodiment, the first antigen binding portion and the second antigen binding portion are each an scFv molecule. In a specific embodiment, the first effector moiety shares an amine-terminal peptide bond or a carboxy-terminal peptide bond with the first antigen-binding moiety, and the second antigen-binding moiety is conjugated to the first effector moiety or 11) the first antigen-binding moiety Amino terminal peptide bonds or carboxy terminal peptide bonds are shared. In a specific embodiment, the immunoconjugate consists essentially of a first single chain effector moiety and a first antigen binding moiety and a second antigen binding moiety. In an even more specific embodiment, the first antigen binding portion and the second antigen binding portion are each a Fab molecule. In one embodiment, the first effector moiety shares a carboxy terminal peptide bond with the first antigen binding moiety and further shares an amine terminal peptide bond with the second antigen binding moiety. In another embodiment, the first antigen binding portion shares a carboxy terminal peptide bond with the first effector moiety (especially the single chain effector moiety) and further shares an amine terminal peptide bond with the second antigen binding moiety. In another embodiment, the first antigen-binding portion shares an amine-terminal peptide bond with the first effector moiety (especially the single-stranded effector moiety), and further shares a few terminal ends with the second antigen-binding moiety. In one embodiment, the 'effector moiety (especially the single bond effector moiety) shares the thiol terminal peptide bond with the 161561.doc • 36-201247221 one heavy bond variable region and further shares the amine terminal with the second heavy chain variable region Peptide bond. In another embodiment, the effector moiety (especially the single chain effector moiety) shares a carboxy terminal peptide bond with the first light chain variable region and further shares an amine terminal peptide with the second light chain variable region. In another embodiment, the 'first heavy or light chain variable region is linked to the first effector moiety (especially the single chain effector moiety) by a carboxy terminal peptide bond, and further linked to the second heavy by an amine terminal peptide bond Key or light key variable area. In another embodiment, the first heavy or light chain variable region is linked to the first effector moiety (especially the single chain effector moiety) by an amino terminal peptide bond, and further linked to the second heavy by a carboxy terminal peptide bond Chain or light chain variable region. In a particular embodiment the 'effector moiety (especially the single chain effector moiety) shares a carboxy terminal peptide bond with the first Fab heavy or light bond and further shares an amine terminal peptide bond with the second Fab heavy or light chain. In another embodiment, the first Fab double or light bond shares a carboxy terminal peptide bond with the first single chain effect moiety and further shares an amine terminal peptide bond with the second Fab heavy or light chain. In other embodiments, the first Fab heavy or light chain shares an amino terminal peptide bond with the first single chain effect moiety and further shares a carboxyl terminal peptide bond with the second Fab heavy or light chain. In one embodiment, the immunoconjugate comprises at least one first effector moiety that shares an amino terminal peptide bond with one or more scFv molecules, and wherein the first effector moiety further shares a carboxy terminal peptide with one or more scFv molecules key. In a particular embodiment, the effector moiety is a single chain effector moiety. In another embodiment, the immunoconjugate comprises at least one first effector moiety (particularly a single-stranded effector moiety), and a first antigen-binding moiety and a second 161561.doc-37-201247221 prion binding moiety, wherein each antigen The binding moiety comprises an scFv molecule conjugated to a constant region comprising an immunoglobulin constant domain at a carboxy-terminal amino acid, and wherein the first antigen-binding moiety is conjugated to the first effector moiety in its strange region An amino terminal amino acid, and wherein the first antigen binding portion and the second antigen binding portion are covalently linked via at least one disulfide bond. In a particular embodiment, the constant region is independently selected from the group consisting of IgG CH1.

IgG CH2, IgG CH3, IgG CK, IgG CjIgE CH4 domain group. In one embodiment, the immunoglobulin domain of the first antigen binding portion is covalently linked to the immunoglobulin domain of the second antigen binding portion via a disulfide bond. In one embodiment, at least one disulfide bond is located at the carboxy terminus of the immunoglobulin domain of the first antigen binding portion and the second antigen binding portion. In another embodiment, at least one disulfide bond is located at the amine end of the immunoglobulin domain of the first antigen binding portion and the second antigen binding portion. In another embodiment, at least two disulfide bonds are located at the amine terminus of the immunoglobulin domain of the first antigen binding portion and the second antigen binding portion. In a particular embodiment, the immunoconjugate comprises a first antigen binding portion and a second antigen binding portion, each antigen binding portion comprising a scFv molecule having a carboxy terminal amino acid conjugated to a constant region comprising an IgG CH1 domain, wherein the An antigen-binding moiety is bonded to an amino-terminal amino acid of a first effect moiety (especially a single-stranded effect moiety) at a carboxy terminal amino acid of a constant region thereof, and wherein the first antigen binding moiety and the second antigen binding moiety are via at least A disulfide bond is covalently linked. The second antigen binding portion of the immunoconjugate can be further joined to the amino terminal amino acid of the second effect moiety at its carboxy terminal amino acid. In one embodiment, the second effect portion is a single chain effect portion. 161561.doc •38- 201247221 In a specific embodiment, the immunoconjugate comprises a first antigen binding portion and a second antigen binding portion, each antigen binding portion comprising a carboxy terminal amino acid linked to a constant comprising an IgG CK domain a scFv molecule of the region, wherein the first antigen-binding portion is bonded to the amino-terminal amino acid of the first effector moiety (especially the single-stranded effector moiety) at a carboxy terminal amino acid of its constant region, and wherein the first antigen-binding moiety And the second antigen binding moiety is covalently linked via at least one disulfide bond. The second antigen-binding portion of the immunoconjugate can be further joined to the amino-terminal amino acid of the second effector moiety at its slow-base amino acid. In one embodiment, the second effect moiety is a single chain effect moiety. In another specific embodiment the 'immunoconjugate comprises a first antigen binding portion and a second antigen binding portion' each antigen binding portion comprising an scFv molecule ligated to a constant region comprising an IgE CH4 domain at a carboxy terminal amino acid, wherein The first antigen-binding portion is joined to the amino-terminal amino acid of the first effector moiety (especially the single-bond effector moiety) at the carboxy terminal amino acid of its constant region and wherein the first antigen-binding moiety and the second antigen-binding portion Partially linked via at least one disulfide bond. The second antigen binding portion of the immunoconjugate can be further joined to the amino terminal amino acid of the second effect moiety at its carboxy terminal amino acid. In one embodiment, the second effector moiety is a single-stranded effector moiety. In another specific embodiment, the immunoconjugate comprises a first antigen binding portion and a second antigen binding portion, each antigen binding portion comprising an scFv molecule ligated to the IgE CH3 domain at a thiol terminal amino acid, wherein the The p-priming moiety is joined to the amino-terminal amino acid of the first effector moiety (especially the single-stranded effector moiety) at its carboxy-terminal amino acid, and wherein the first antigen 161561.doc •39-201247221 binds to the moiety The second antigen binding moiety is covalently linked via at least one disulfide bond. The second antigen binding portion of the immunoconjugate can be further joined to the amino terminal amino acid of the second effect moiety at its carboxyl terminal amino acid. In one embodiment, the second effect portion is a single chain effect portion. In another embodiment, the immunoconjugate comprises a first effector moiety and a second effector moiety, and a first antigen binding moiety and a second antigen binding moiety, wherein each antigen binding moiety comprises a heavy chain or a light chain carboxyl terminal amine group The acid is bonded to the Fab molecule of the IgG1 CH3 domain, and wherein each IgG1 CH3 domain is joined to an amino-terminal amino acid of an effector moiety at its respective carboxy terminal amino acid, and wherein the first antigen-binding moiety and the second The antigen binding moiety is covalently linked via at least one disulfide bond. In a particular embodiment, the first effector moiety and/or the second effector moiety are single-stranded effect moieties. In another embodiment, the IgGl CH3 domain of the antigen-binding moiety can be joined by a disulfide bond. In another embodiment, at least one disulfide bond is located at the carboxy terminus of the IgG 1 CH3 domain of the first antigen binding portion and the second antigen binding portion. In another embodiment, at least one disulfide bond is located at the amino terminus of the IgGl CH3 domain of the first antigen binding portion and the second antigen binding portion. In another embodiment, at least two disulfide bonds are located at the amino terminus of the IgGl CH3 domain of the first antigen binding portion and the second antigen binding portion. In some embodiments' immunoconjugates comprise one or more proteolytic cleavage sites between the effector moiety and the antigen binding moiety. The components of the immunoconjugate (eg, the antigen binding portion and/or the effector portion) can be directly or via various linkers as described herein or known in the art (especially comprising one or more amino acids, typically about 2 Linked to a peptide linker of 20 amino acids). Suitable 161561.doc -40- 201247221 Non-immunogenic linker peptides include, for example, (G4S)n, (SG4)n4 04(804)11 linker peptides, wherein η is generally a number from 1 to 10, usually from 2 to 4 . Antigen-binding portion The antigen-binding portion of the immunoconjugate of the invention is generally a polymorphic molecule that binds to a particular antigenic determinant and is capable of directing the entity to which it is attached (eg, an effector moiety or a second antigen-binding moiety) to a target site For example, a particular type of tumor cell or tumor stroma with the antigenic determinant. The immunoconjugate can bind to, for example, an antigenic determinant found on the surface of tumor cells, on the surface of virus-infected cells, on the surface of other diseased cells, in serum and/or in extracellular matrix (ECM). Non-limiting examples of tumor antigens include MAGE, MART-1/Melan-A, gplOO, dipeptidyl peptidase IV (DPPIV), adenosine deaminase binding protein (ADAbp), cyclophilin b, colorectal associated antigen (CRC)-C017-1 A/GA733, carcinoembryonic antigen (CEA) and its immunogenic epitopes CAP-1 and CAP-2, etv6, ami 1, prostate specific antigen (PSA) and their immunogenicity Antigenic epitopes PSA-1, PSA-2 and PSA-3, prostate specific membrane antigen (PSMA), T cell receptor/CD3-$ chain, MAGE family tumor antigen 〇, jWMAGE-Al, MAGE-A2, MAGE- A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-All, MAGE-A12, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE-C1, MAGE-C2, MAGE-C3, MAGE-C4, MAGE-C5), GAGE family tumor antigen (^'] WGAGE-1, GAGE- 2. GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, GAGE-9), 161561.doc 41 - 201247221 BAGE, RAGE, LAGE-1, NAG, GnT-V , MUM-1, CDK4, tyrosinase, p53, MUC family, HER2/neu, p21ras, RCAS1, a-fetoprotein, E-cadherin , a-sodium hydrocarbon, β-sodium hydrocyclone and γ-soda, pl20ctn, gpl00Pmelll7, PRAME, NY-ESO-1, cdc27, colon adenomatous polyposis protein (APC), lining protein (fodrin) , connexin 37, Ig-individual genotype, pl5, gp75, GM2 and GD2 gangliosides, viral products (such as human papillomavirus protein), Smad family tumor antigens, lmp-1, P1A, EBV-encoded nuclei Antigen (EBNA)-1, brain glycogen phosphorylase, SSX-1, SSX-2 (HOM-MEL-40), SSX-1, SSX-4, SSX-5, SCP-1 and CT-7, and Non-limiting examples of c-erbB-2 »viral antigens include influenza virus hemagglutinin, Epstein-Barr virus LMP-1, hepatitis C virus E2 glycoprotein, HIV gpl60, and HIV gpl20. Non-limiting examples of ECM antigens include syndecan, heparinase, integrin, osteopontin, connectin, good adhesion protein, laminin, laminin EGF, lectin, fibronectin, notch, tenascin, and matrix metalloproteinase. The immunoconjugates of the invention may bind to specific non-limiting examples of cell surface antigens: FAP, Her2, EGFR, IGF-1R, CD2 (T cell surface antigen), CD3 (heteromultimers associated with TCR), CD22 (B cell receptor), CD23 (low affinity IgE receptor), CD25 (IL-2 receptor a chain), CD30 (cytokine receptor), CD33 (myeloid cell surface antigen), CD40 (tumor necrosis factor receptor) , IL-6R (IL6 receptor), CD20, MCSP, c-Met, protein containing CUB domain (CDCP1) and 161561.doc • 42·201247221 PDGFpR (p platelet-derived growth factor receptor). In one embodiment, the antigen binding portion is directed against an antigen presented on a tumor cell or in a tumor cell environment. In the embodiment, the antigen junction β knife line is directed to an A1 domain (TNC Α1) selected from the group consisting of fibroblast-activated protein (FAp) and tenosin c (tendon protein (: octa 2 domain (TNC) Α2), an antigen of a group of fibronectin-extradomain B (EDB), carcinoembryonic antigen (CEA), and melanoma-associated chondroitin proteoglycan (Mcsp). In one embodiment, the immunoconjugate of the present invention comprises Two or more antigen-binding portions, wherein each of the antigen-binding portions specifically binds to the same antigenic determinant. In another embodiment, the immunoconjugate of the invention comprises two or more antigen-binding portions, wherein Each of the antigen binding portions specifically binds to a different antigenic determinant. The antigen binding portion may be any type of antibody or fragment thereof that retains specific binding to an antigenic determinant. In one embodiment, the antigen binding portion is an antibody or antibody. Fragments. Antibody fragments include, but are not limited to, vH fragments, VL fragments, Fab fragments, F(ab')2 fragments, scFv fragments, Fv fragments, minibodies, bifunctional antibodies, trifunctional antibodies, and Functional antibodies (see for example

Hudson and Souriau, Nature Med 9, 129-134 (2003)). Particularly suitable antibody fragments are Fab fragments and scFv fragments. Thus, in one embodiment the 'antigen binding portion' is selected from the group consisting of a Fab molecule and an scFv molecule. In one embodiment, the antigen binding portion is a Fab molecule. In another embodiment, the antigen binding portion is a scFv molecule. In one embodiment, the immunoconjugate comprises at least one, usually two or more, fibronectin outer domain B (EDB) specific antigen binding portions 161561.doc • 43·201247221 points. In another embodiment, the immunoconjugate comprises at least one, usually two or more, antigen binding portions that compete with the monoclonal antibody L1 9 for binding to the antigenic epitope of EDB. See, for example, PCT Publication WO 2007/128563 A1, which is incorporated herein by reference in its entirety. In another embodiment, the 'immunoconjugate comprises a polypeptide sequence' wherein the first Fab heavy chain derived from the L19 monoclonal antibody shares a carboxy terminal peptide bond with the IL-2 molecule, and the IL-2 molecule is derived from the L19 single The second Fab heavy chain of the strain antibody shares a carboxy terminal peptide bond. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the first Fab heavy bond derived from the L19 monoclonal antibody shares a carboxy terminal peptide bond with the IL-12 molecule and the IL-12 molecule is derived from the L19 single The second Fab heavy chain of the strain antibody shares a carboxy terminal peptide bond. In another embodiment, the immunoconjugate comprises a polypeptide sequence wherein the first Fab heavy bond derived from the L19 monoclonal antibody shares a carboxy terminal peptide bond with the IFNα molecule, and the IFN α molecule is further derived from the L19 monoclonal antibody The second Fab heavy chain shares a tick-terminal peptide bond. In another embodiment, the immunoconjugate comprises a polymorphic sequence, wherein the first Fab heavy bond derived from the L丨9 monoclonal antibody shares a carboxy terminal peptide bond with the GM-CSF molecule, and the GM-CSF molecule is further derived from the source The second Fab heavy bond of the Li9 monoclonal antibody shares an antagonistic terminal peptide bond. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the first (7) derived from the L19 monoclonal antibody shares a carboxy terminal peptide bond with the 匕^ molecule, and the IL-2 molecule is further derived from the L19 monoclonal antibody. The second scFv shares a slow-acting terminal peptide bond β. In a more specific embodiment, the immunoconjugate comprises a polymorphic sequence of SEQ ID NO: 91 or a variant thereof that retains functionality. In another embodiment, the immunoconjugate comprises a Fab light chain derived from a l19 monoclonal antibody. In a more specific embodiment, the immunoconjugate 161561.doc 44-201247221 comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or SEQ ID NO: 92 100% of the polypeptide sequence or a variant thereof that retains functionality. In yet another embodiment, the immunoconjugate comprises two of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% with SEQ ID NO: 91 and SEQ ID NO: 92. Or 100% of the polypeptide sequence or a variant thereof that retains functionality. In a more specific embodiment, the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide of SEQ ID NO: 98 A sequence or a variant thereof that retains functionality. In still another embodiment, the immunoconjugate comprises two at least about 80%, 85%, 90%, 95%, 96%, 97 of SEQ ID NO: 98 and SEQ ID NO: 92. /〇, 98%, 99%, or 100% of the polypeptide sequence or a variant thereof that retains functionality. In a more specific embodiment, the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% '99% or 100% of the polypeptide of SEQ ID NO: 99 A sequence or a variant thereof that retains functionality. In yet another embodiment, the immunoconjugate comprises two of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% of SEQ ID NO: 99 and SEQ ID NO: 92. Or a 100% identical polypeptide sequence or a variant that retains its functionality. In a more specific embodiment, the immunoconjugate comprises at least about 80%, 85%, 90 of SEQ ID NO: 100. /. 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In yet another embodiment, the immunoconjugate comprises two at least about 80%, 85% '90% '95%, 96%, 97%, 98% > 99 with SEQ ID NO: 100 and SEQ ID NO: % or 100% of the polypeptide sequence or a variant thereof that retains functionality. In a particular embodiment of -45-161561.doc 201247221, the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% of SEQ ID NO: 101 Or a 100% identical polypeptide sequence or a variant that retains its functionality. In yet another embodiment, the immunoconjugate comprises two at least about 80%, 85%, 90%, 95%, 960/〇, 97%, 98%, 99 with SEQ ID NO: 101 and SEQ ID NO: % or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the polypeptide is covalently linked by, for example, a disulfide bond. In one embodiment, an immunoconjugate of the invention comprises at least one, usually two or more, tenosin A1 domain (TNC-A1) specific antigen binding portions. In another embodiment, the immunoconjugate comprises at least one, usually two or more, antigen binding portions that compete with the monoclonal antibody F16 for binding to the epitope of TNC-A1. See, for example, PCT Publication WO 2007/1285 63 A1, which is incorporated herein by reference in its entirety. In one embodiment, the immunoconjugate comprises at least one, typically two or more, myipein A1 domain and/or A4 domain (TNC-A1 or TNC-A4 or TNC_A1/A4) specific antigen binding moiety. In another embodiment, the immunoconjugate comprises a polymorphic sequence wherein the myosin A1 domain-specific first Fab heavy bond shares a carboxy terminus with an IL-2 molecule, an IL-12 molecule, an IFN alpha molecule, or a GM-CSF molecule The cleavage bond, while the IL-2 molecule, IL-12 molecule, IFN alpha molecule or GM-CSF molecule, in turn, shares a carboxy terminal peptide bond with the tendon peptone domain specific second Fab heavy chain. In another embodiment, the immunoconjugate comprises a polypeptide sequence wherein the tenosin A1 domain-specific first Fab heavy chain shares a carboxy terminal peptide bond with the IL-2 molecule, and the IL-2 molecule is further associated with the tenosin A1 domain The specific second Fab heavy chain shares a carboxy terminal peptide bond. In another embodiment, the immunization 161561.doc • 46-201247221 conjugate comprises a polypeptide sequence wherein the tenosin A1 domain specific first scFv shares a carboxy terminal peptide bond with the IL-2 molecule, and the IL-2 molecule A tick-terminal peptide bond is shared with a tenosin A1 domain-specific second scFv. In a specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% of SEQ ID NO: 8 or SEQ ID NO: 99% or 100% of the heavy chain variable region sequences or variants thereof retaining functionality. In another specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% of SEQ ID NO: 6 or SEQ ID NO: 7. , 99% or 100% identical light chain variable region sequences or variants thereof that retain functionality. In a more specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% of SEQ ID NO: 8 or SEQ ID NO: a 99% or 100% heavy chain variable region sequence or a variant thereof which retains functionality, and at least about 80%, 85%, 90%, 95% with SEQ ID NO: 6 or SEQ ID NO: 7. '960/〇, 970/〇, 98%, 99%, or 100%—to the light chain variable region sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide of SEQ ID NO: A sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99 of SEQ ID NO: 96 or SEQ ID NO: 105 % or 100% of the polypeptide sequence or variant thereof retaining functionality" In another specific embodiment, the immunoconjugate of the invention comprises at least about 80% of SEQ ID NO: 97 or SEQ ID NO: 115, • 47· 161561.doc 201247221 85〇/〇, 90%, 95〇/〇, 96% '97%, 98%, 99% or 100%—the polypeptide sequence or a variant of its retention functionality In a more specific embodiment, the immunoconjugate of the invention comprises two at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% of SEQ ID NO: 96 and SEQ ID NO: 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises two at least about 80%, 85%, 90%, 95%, 96%, 97%, 98 with SEQ ID NO: 105 and SEQ ID NO: %, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In one embodiment, the immunoconjugate comprises at least one, typically two or more, Tendin A2 domain (TNC-A2) specific antigen binding portions. In another embodiment, the immunoconjugate comprises a polypeptide sequence wherein the tenosin A2 domain-specific first Fab heavy chain shares a carboxy terminal peptide with an IL-2 molecule, an IL-12 molecule, an IFN alpha molecule, or a GM-CSF molecule The bond, while the IL-2 molecule, IL-12 molecule, IFN alpha molecule or GM-CSF molecule, in turn, shares a carboxy terminal peptide bond with the tendon peptone 2 domain-specific second Fab heavy chain. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the tenosin A2-domain specific first Fab heavy chain shares a carboxy terminal peptide bond with the IL-2 molecule, and the IL-2 molecule is specifically specific to the tendon A2 domain The second Fab heavy chain shares a carboxy terminal peptide bond. In a specific embodiment, the antigen binding portion of the immunoconjugate comprises and is selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 71, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83 and SEQ ID NO: 85. Sequence 歹 1J at least about 80%, 85% '90% ' 95% ' 96% ' 97% ' 98% &gt 161561.doc -48- 201247221 99% or 100% of the heavy chain variable region sequence or a variant thereof that retains functionality. In another specific embodiment, the antigen binding portion of the immunoconjugate comprises and is selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ The sequence of ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, and SEQ ID NO: 84 is at least about 80%, 85%, 90%, 95%, 96%, 97 %, 98%, 99% or 100% of the light chain variable region sequences or variants thereof retaining functionality. In a more specific embodiment, the antigen binding portion of the immunoconjugate comprises and is selected from the group consisting of SEQ ID NO: 5, SEQ ID NO:

71. The sequence of the population of SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, and SEQ ID NO: 85 is at least about 80 %, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the heavy chain variable region sequence or a variant thereof which retains functionality, and is selected from the group consisting of SEQ ID NO : SEQ ID NO: 4, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82 And the sequence of the population of SEQ ID NO: 84 is at least about 80%, 85%, 90%, 95%, 960/〇, 97%, 98%, 99% or 100% of the light chain variable region sequence or Functional variants are retained. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80%, 85%, 90% of the sequence selected from the group consisting of SEQ ID NO: 117, SEQ ID NO: 11 8 and SEQ ID NO: 119 , 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80% of a sequence selected from the group consisting of SEQ ID 49 161561.doc 201247221 NO: 120, SEQ ID NO: 121, and SEQ ID NO: 122 , 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In a more specific embodiment, the immunoconjugate of the invention comprises at least about 80%, 85%, 90% of the sequence selected from the group consisting of SEQ ID NO: 117, SEQ ID NO: 118, and SEQ ID NO: 119 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof which retains functionality, and is selected from the group consisting of SEQ ID NO: 120, SEQ ID NO: 121 and SEQ ID The sequence of NO: 122 is at least about 80%, 85°/. , 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises two at least about 80%, 85%, 90%, 95%, 96 of SEQ ID NO: 117 and SEQ ID NO: 121 or SEQ ID NO: 122 %, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises two at least about 80%, 85%, 90%, 95%, 96 of SEQ ID NO: 118 and SEQ ID NO: 120 or SEQ ID NO: 121 %, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises two at least about 80%, 85%, 90%, 95%, 96%, 97%, 98 with SEQ ID NO: 119 and SEQ ID NO: %, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In one embodiment, the immunoconjugate comprises at least one, usually two or more fibroblast activated protein (FAP) specific antigen binding portions 161561.doc -50· 201247221

Minute. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the FAP-specific first Fab heavy chain shares a carboxy terminal peptide bond with an IL-2 molecule, an IL-12 molecule, an IFN alpha molecule, or a GM-CSF molecule, and IL The -2 molecule, IL-12 molecule, IFN alpha molecule or GM-CSF molecule in turn shares a carboxy terminal peptide bond with the FAP-specific second Fab heavy chain. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the FAP-specific first Fab heavy chain shares a carboxy terminal peptide bond with the IL-2 molecule, and the IL-2 molecule is in turn shared with the FAP-specific second Fab heavy chain Carboxy terminal peptide bond. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the FAP-specific first Fab heavy chain shares a carboxy terminal peptide bond with the IL-12 molecule, and the IL-12 molecule is in turn shared with the FAP-specific second Fab heavy chain Carboxy terminal peptide bond. In a specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or a sequence selected from the group consisting of 100% of the heavy chain variable region sequence or variant thereof retaining functionality: SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19. SEQ ID NO: 21, SEQ ID NO: 23, SEQ

ID NO: 25 ' SEQ ID NO: 27 > SEQ ID NO: 29 ' SEQ ID

NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ

ID NO: 51 'SEQ ID NO: 53 ' SEQ ID NO: 55 ' SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO : 67 and SEQ ID NO: 69. In another specific embodiment, the antigen binding portion of the immunoconjugate comprises 161561.doc • 51 - 201247221

The sequence of the population of the following composition is selected to be at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the light chain variable region sequence or its retention functionality Variants: SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66 and SEQ ID NO: 68. In a more specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99° to a sequence selected from the group consisting of /. Or a 100% heavy chain variable region sequence or a variant thereof that retains functionality: SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19 SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO : 53. SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ 161561.doc -52- 201247221 ID NO: 67 and SEQ ID NO: 69, and at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the sequence selected from the group consisting of: Consistent light chain variable region sequences or variants thereof retaining functionality: SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36 SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, and SEQ ID NO: 68. In another specific embodiment, the immunoconjugate of the invention comprises and is selected from the group consisting of SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID The sequence of NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, and SEQ ID NO: 111 is at least about 80%, 85%, 90%, 95°/〇, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80% of a sequence selected from the group consisting of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, and SEQ ID NO: 116, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In a more specific embodiment, the immunoconjugate of the invention comprises at least about 80 SEQ ID NO: SEQ ID NO: 103, 161561.doc • 53-201247221 SEQ ID NO: 107 and SEQ ID NO: 108 /〇, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical polypeptide sequence or a variant thereof that retains functionality, and at least about 80% of SEQ ID NO: 113 , 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80% of a sequence selected from the group consisting of SEQ ID NO: 102, SEQ ID NO: 109, SEQ ID NO: 110, and SEQ ID NO: 111, 85%, 90%, 95%, 96%, 97°/〇, 98%, 99%, or 100% of the polypeptide sequence or a variant thereof that retains functionality, and at least about 80% of SEQ ID NO: 112 , 85%, 90%, 950/〇, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises two at least about 80%, 85%, 90%, 95%, 96%, 97%, 98 with SEQ ID NO: 104 and SEQ ID NO: %, 99% or 100% of the polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises two and eight £(^1〇1^〇: 106 and 8 £(5 1〇)^:116 at least about 80%, 85%, 90 %, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof retaining functionality. In another specific embodiment, the immunoconjugate of the invention comprises two At least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or its retention functionality with SEQ ID NO: 108 and SEQ ID NO: 113 In another particular embodiment, the immunoconjugate of the invention comprises two at least about 80%, 161561.doc -54 - 201247221 85%, 90% with SEQ ID NO: 109 and SEQ ID NO: 112 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof which retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises two SEQ ID NO: 11 and SEQ ID NO: 112 at least about 80%, 85%, 90%, 95%, 96%, 97°/〇, 98%, 99% or 100% of the polypeptide sequence or its retention Functional variant. In another specific embodiment, the immune knot of the invention The composition comprises two at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% of SEQ ID NO: 111 and SEQ ID NO: 112 A polypeptide sequence or a variant thereof that retains functionality. In one embodiment, the immunoconjugate comprises at least one, typically two or more melanoma chondroitin sulfate proteoglycan (MCSP) specific antigen binding portions. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the MCSP-specific first Fab heavy chain shares a carboxy terminal peptide bond with an IL-2 molecule, an IL-12 molecule, an IFN alpha molecule, or a GM-CSF molecule, and IL- The 2 molecule, IL-12 molecule, IFN alpha molecule or GM-CSF molecule in turn shares a carboxy terminal peptide bond with the MCSP-specific second Fab heavy chain. In another embodiment, the immunoconjugate comprises a polypeptide sequence wherein MCSP specificity The first Fab heavy chain shares a carboxy terminal peptide bond with the IL-2 molecule, and the IL-2 molecule in turn shares a carboxy terminal peptide bond with the MCSP-specific second Fab heavy chain. In a particular embodiment, the antigen binding of the immunoconjugate Partially comprising at least about 80%, 85%, 90%, 95 of the sequence of SEQ ID NO: 86 or SEQ ID NO: 88 %, 96%, 97%, 98%, 99% or 100% of the heavy chain variable region sequences or variants thereof retaining functionality. In another specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85% '90%, 95% of the sequence 161561.doc-55-201247221 of SEQ ID NO: 87 or SEQ ID NO: 90 , 96%, 97%, 98%, 99%, or 100% of the light chain variable region sequence or a variant thereof that retains functionality. In a more specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97% of the sequence of SEQ ID NO: 86 or SEQ ID NO: 88, 98%, 99% or 100% of the heavy chain variable region sequence or a variant thereof retaining functionality, and at least about 80%, 85%, 90 of the sequence of SEQ ID NO: 87 or SEQ ID NO: 90 %, 9"/, 96%, 97%, 98%, 99%, or 100% of the light chain variable region sequence or a variant thereof that retains functionality. In a more specific embodiment, the immunoconjugate The antigen binding portion comprises at least about 80%, 85% '90% '95% '96%, 97%, 98% '99% or 100% of the sequence of SEQ ID NO: 86 And at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the sequence of SEQ ID NO: 87, resulting in a light chain variable region sequence. In a particular embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80 〇/〇, 85%, 90%, 95%, 96%, 97%, 98 〇/〇, 99 of the sequence of SEQ ID NO:88. % or 100% of the heavy chain variable region sequence, and at least about 80% of the sequence of SEQ ID NO: 87 , 85%, 90% '95%, 96%, 97%, 980/〇, 99% or 100% of the light chain variable region sequence. In another specific embodiment, the immunoconjugate of the invention comprises At least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or its retention functionality with SEQ ID NO: 123 or SEQ ID NO: 125 A variant of the invention. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80%, 85%, 90%, 161561.doc-56-201247221 95 with SEQ ID NO: 124 or SEQ ID NO: %, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or a variant thereof which retains functionality. In a more specific embodiment, the immunoconjugate of the invention comprises SEQ ID NO: 123 or SEQ ID NO: 125 is at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical polypeptide sequence or a variant thereof which retains functionality, and SEQ ID NO: 124 or SEQ ID NO: 127 at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of the polypeptide sequence or its retention functionality Variant. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of SEQ ID NO: 123 - a polypeptide sequence or a variant thereof that retains functionality, and at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of SEQ ID NO: 124 A polypeptide sequence or a variant thereof that retains functionality. In another specific embodiment, the immunoconjugate of the invention comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% of SEQ ID NO: a consensus polypeptide sequence or a variant thereof that retains functionality, and at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100 with SEQ ID NO: 124 % - a polypeptide sequence or a variant thereof that retains functionality. In one embodiment, the immunoconjugate comprises at least one, usually two or more carcinoembryonic antigen (CEA) specific antigen binding portions. In another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein CEA is specific The first Fab heavy chain shares a carboxy terminal peptide bond with an IL-2 molecule, an IL-12 molecule, an IFNα molecule or a GM-CSF molecule, while the IL-2 molecule, IL-12 molecule, IFNα 161561.doc •57· The 201247221 molecule or GM-CSF molecule in turn shares a carboxy terminal peptide bond with a CEA-specific second Fab heavy chain. In still another embodiment, the immunoconjugate comprises a polypeptide sequence, wherein the CEA-specific first Fab heavy chain shares a carboxy-terminal peptide bond with the IL-2 molecule, and the IL-2 molecule shares a carboxy terminus with the CEA-specific second Fab heavy chain Peptide bond. In a specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100 of the sequence of SEQ ID NO: 154 % - a heavy chain variable region sequence or a variant thereof that retains functionality. In another specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80 〇/〇, 85% '90% ' 95% ' 96% ' 97%, 98%, 99 with the sequence of SEQ ID NO: 155 % or 100% of the light chain variable region sequence or a variant thereof that retains functionality. In a more specific embodiment, the antigen binding portion of the immunoconjugate comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or the sequence of SEQ ID NO: 154 or 100% of the heavy chain variable region sequence or a variant thereof retaining functionality, and at least about 80 Å/〇, 85% > 90% > 95%, 96%, of the sequence of SEQ ID NO: 155 97%, 98% '99% or 100%--the light chain variable region sequence or a variant thereof that retains functionality. The antigen binding portion of the invention comprises at least about 80%, 85%, 90%, 95%, 96%, 97%, 98 comprising the sequences described in SEQ ID Nos. 3 to 127, including functional fragments or variants thereof. %, 99% or 100% of the antigen binding portion of the sequence. The present invention also encompasses antigen-binding portions comprising the sequences of SEQ ID Nos. 3 to 127 having conservative amino acid substitutions. It is understood that in the sequences SEQ ID No 91, 93, 94, 95, 96, r〇f 103, 104 , 105, 106, 108, 109, 110, 111, 117, 118, 119, 161561.doc .58 - 201247221 123 and 125 'Human ratio-2 sequence (see SEQ ID NO: 1) can be replaced with IL-2 Analog sequences, especially the mutant IL2 described herein (see seqID NO: 2). Effector Part of the Immunoconjugate The effector moiety used in the present invention is generally a polypeptide which affects cellular activity, for example, via a signal transduction pathway. Thus, the effector portion of an immunoconjugate suitable for use in the present invention can be associated with a member-mediated signaling that transmits signals from outside the cell membrane to modulate intracellular responses. For example, the effect of the immunoconjugate may be in part a cytokine. In a particular embodiment, the effect moiety is a single chain effect moiety as defined herein. In one embodiment, one or more effector portions (usually single-stranded effect portions) of the immunoconjugate of the invention are cytokines selected from the group consisting of IL-2, GM-CSF, IFN-[alpha], and IL_ 12. In one embodiment, the effector moiety is IL-2. In another embodiment, the one or more single bond effect portions of the immunoconjugate are cytokines selected from the group consisting of IL_8, MIP-ia, MIP-Ιβ, and TGF-β. In one embodiment, the effector portion of the immunoconjugate, particularly the single-chain effector moiety, is IL-2. In a particular embodiment, the IL_2 effector moiety can cause one or more cellular responses selected from the group consisting of: activation of T lymphocyte proliferation, activation of tau lymphocyte differentiation, cytotoxic T cell (CTL) survival, activation of B cells Proliferation, activation of b cell differentiation, natural killer (Νκ) cell proliferation, NK cell differentiation, activation of τ cell or ΝΚ cell-derived cytokine secretion and ΝΚ/lymphocyte activation killer (LAK) anti-tumor cytotoxicity. In certain embodiments, the 'IL-2 effector moiety is a mutant IL-2 effector moiety that comprises at least one reduced or eliminated protuberance compared to the uninduced IL-2 effector moiety. 161561.doc • 59· 201247221 variable IL- 2 The effector moiety has affinity for the alpha subunit of the IL-2 receptor (also known as CD25) but retains the mutant IL-2 effector moiety for the intermediate affinity IL-2 receptor (by the beta and gamma subunits of the IL-2 receptor) The amino acid mutation of the affinity of the composition). In one embodiment, the amino acid is mutated to an amino acid substitution. In a specific embodiment, the mutant IL-2 effector moiety comprises 1, 2 or at a position selected from the group consisting of residues 42, residues 45 and residues 72 corresponding to human IL-2. Three amino acid substitutions. In a more specific embodiment, the mutant IL-2 effector moiety comprises three amino acid substitutions at positions corresponding to residues 42, residues 45 and residues 72 of human IL-2. In an even more specific embodiment, the mutant IL-2 effect portion is human IL-2 comprising an amino acid substituted for F42A, Υ45Α and L72G. In one embodiment, the mutant IL-2 effector portion additionally comprises an amino acid mutation at a position corresponding to position 3 of human IL-2, which mutation abolishes the 0-glycosylation site of IL-2. This other amino acid mutation is especially an amino acid substitution in which a threonine residue is replaced with an alanine residue. The sequence comprising the amino acid-substituted tetraploid mutation (QM) IL-2 of Τ3 A, F42A, Y45A and L72G is shown in SEQ ID NO: 2. Suitable mutant IL-2 molecules are described in more detail in European Patent Application No. EP11153964.9. Mutant IL-2 molecules useful as part of the effector in an immunoconjugate can be prepared by deletion, substitution, insertion or modification using genetic or chemical methods well known in the art. Genetic methods can include site-specific mutation induction encoding DNA sequences, PCR, gene synthesis, and the like. The correct nucleotide change is verified by, for example, sequencing. In this regard, the nucleotide sequence of native IL-2 has been described by Taniguchi et al. (Nature 302, 305-10 (1983)) and the nucleic acid encoding human IL-2 is available from, for example, the American Type Culture Collection Center 16156I.doc. -60- 201247221 (American Type Culture Collection) (Rockville MD) public depository. An exemplary sequence of human IL-2 is shown in SEQ ID NO: 1. Substitution or insertion may include both natural as well as non-natural amino acid residues. Amino acid modifications include well-known chemical modification methods such as addition or removal of glycosylation sites or carbohydrate linkages, and the like. In one embodiment, the effector portion of the immunoconjugate, particularly the single-chain effector moiety, is GM-CSF. In a particular embodiment, the GM-CSF effector moiety can initiate proliferation and/or differentiation of granulocytes, monocytes or dendritic cells. In one embodiment, the effector portion of the immunoconjugate, particularly the single-button effect moiety, is IFN-[alpha]. In a particular embodiment, the iFN-alpha effector moiety elicits one or more cellular responses selected from the group consisting of viral replication in a virus-infected cell and up-regulation of a major histocompatibility complex I (MHC I). In another specific embodiment, the IFN-a effector moiety inhibits tumor cell proliferation. In one embodiment, the effector portion of the immunoconjugate, particularly the single chain effect portion, is IL-12. In a particular embodiment, the IL-12 effector moiety elicits one or more cellular responses selected from the group consisting of NK cell proliferation, NK cell differentiation, T cell proliferation, and tau cell differentiation. In one embodiment, the effector portion of the immunoconjugate, particularly the single chain effector moiety, is IL-8. In a particular embodiment, the IL_8 effector moiety can elicit chemotaxis of neutrophils. In one embodiment, the effector portion of the immunoconjugate, particularly the single-stranded effect portion, is MIp_丨a. In a particular embodiment, the MIp-la effector moiety can elicit chemotaxis of monocytes and tau lymphocytes. In one embodiment, the effect portion of the immunoconjugate, particularly the single-stranded effect, is divided into MIP-Ιβ❶. In a specific embodiment, the MIp_1J3 effector moiety can be 161561.doc-61 - 201247221 to induce monocytes and tau lymphocytes. Chemotaxis. In one embodiment, the effector portion of the immunoconjugate, particularly the single chain effector moiety, is TGF-[beta]. In a particular embodiment, the TGF-beta effector moiety can elicit one or more cellular responses selected from the group consisting of: monocyte chemotaxis, macrophage chemotaxis, activation of IL-1 in macrophages Up-regulation and up-regulation of IgA in activated sputum cells. Antibodies Antibodies suitable for use in the present invention include antibodies or antibody fragments that bind to a particular antigenic determinant (e.g., a particular tumor cell antigen) and comprise an Fc region. In certain embodiments, the anti-system is directed against an antigen presented on a tumor cell. Specific target antigens suitable for use in the antibodies of the invention include antigens expressed on the surface of tumor cells including, but not limited to, cell surface receptors such as epidermal growth factor receptor (EGFR), insulin-like growth factor receptor (IGFR). And platelet-derived growth factor receptor (PDGFR); prostate specific membrane antigen (PSMA), carcinoembryonic antigen (CEA), dipeptidyl peptidase IV (CD26, DPPIV), FAP, HER2/neu, HER-3, E-cadherin, CD20, melanoma-associated chondroitin proteoglycan (MCSP), c-Met, CUB domain-containing protein 1 (CDCP1), and squamous cell carcinoma antigen (SCCA). In a specific embodiment, the anti-system is directed to selected from the group consisting of CD20, epidermal growth factor receptor (EGFR), HER2, HER3, insulin-like growth factor 1 receptor (IGF-1R), carcinoembryonic antigen (CEA), c-Met An antigen comprising a CUB domain-derived protein 1 (CDCP1) and a melanoma-associated chondroitin sulfate proteoglycan (MCSP). In one embodiment, the antibody (multispecific antibody) is 161561.doc-62-201247221 for two or more selected from the group consisting of CD20, epidermal growth factor receptor (EGFR), HER2, HER3, insulin-like growth Antigens of the Factor 1 receptor (jgf_1R), carcinoembryonic antigen (CEA), c-Met, CUB domain-containing protein 1 (CDCP1), and melanoma-associated chondroitin proteoglycan (MCSP). A specific anti-CD20 antibody suitable for use in the present invention is a humanized IgG-type scorpion-type anti-CD20 antibody having the binding specificity of a murine B-Lyl antibody (Poppema and Visser, Biotest Bulletin 3, 131-139 (1987)). Particularly suitable are humanized IgG class II anti-CD20 antibodies comprising: a) CDR1 (SEQ ID NO: 128), CDR2 (SEQ ID NO: 129) and CDR3 (SEQ ID NOM30) in the heavy chain variable domain And b) CDR1 (SEQ ID NO: 131), CDR2 (SEQ ID NO: 132) and CDR3 (SEQ ID NO: 133) in the light chain variable domain. Specifically, the heavy chain variable region framework regions (FR) FR1, FR2 and FR3 of the antibody are human FR sequences encoded by the VH1_10 human germline sequence, and the heavy chain variable region FR4 of the antibody is a JH4 human germline sequence. The human FR sequence encoded, the light chain variable regions FR FR1, FR2 and FR3 of the antibody are human FR sequences encoded by the VK_2_40 human germline sequence, and the light chain variable region FR4 of the antibody is the JK4 human germline sequence The encoded human FR sequence. More specific anti-CD20 antibodies suitable for use in the present invention comprise the heavy chain variable domain SEQ ID NO: 134 and the light chain variable domain SEQ ID NO: 135. Such anti-CD20 antibodies are described in WO 2005/044859, which is incorporated herein by reference in its entirety. 161561.doc • 63· 201247221 A specific anti-EGFR antibody suitable for use in the present invention is a humanized IgG class antibody having the binding specificity of a rat ICR62 antibody (Modjtahedi et al, Br J Cancer 67, 247-253 (1993)) . Particularly suitable are humanized IgG class anti-EGFR antibodies comprising: a) CDR1 (SEQ ID NO: 136), CDR2 (SEQ ID NO: 137) and CDR3 (SEQ ID NO: 138) in the heavy chain variable domain And b) CDR1 (SEQ ID NO: 139), CDR2 (SEQ ID NO: 140) and CDR3 (SEQ ID NO: 141) in the light chain variable domain. More specific anti-EGFR antibodies suitable for use in the present invention comprise a heavy chain variable domain SEQ ID NO: 142 and a light chain variable domain SEQ ID NO: 143. Such anti-EGFR antibodies are described in WO 2006/082515 and WO 2008/017963, each of which is incorporated herein by reference in its entirety. Other suitable humanized IgG anti-EGFR antibodies suitable for use in the present invention include cetuximab/IMC-C225 (Erbitux®, described in Goldstein et al, Clin Cancer Res 1, 1311-1318 (1995)) , panitumumab/ABX-EGF (Vectibix®, described in Yang et al, Cancer Res 59, 1236-1243 (1999), Yang et al, Critical Reviews in Oncology/Hematology 38, 17-23 (2001) )), nimotuzumab/h-R3 (TheraCim®' is described in Mateo et al, Immunotechnology 3, 71-81 (1997); Crombet-Ramos et al, Int J Cancer 101, 567-575 (2002); Boland and Bebb, Expert Opin Biol Ther 9, 1199-1206 (2009)), matuzumab/EMD 72000 (described in Bier et al., Cancer 161S61.doc • 64-201247221

Immunol Immunother 46, 167-173 (1998) 'Kim, Curr Opin Mol Ther 6, 96-103 (2004)) and Zalumumumab / 2F8 (described in Bleeker et al, J Immunol 173, 4699) -4707 (2004) 'Lammerts van Bueren, PNAS 105, 6109-6114 (2008)). Specific anti-IGF-1R antibodies suitable for use in the present invention are described in WO 2005/005635 and WO 2008/077546, the contents of each of each of each of each of IGF-1) and insulin-like growth factor-2 (IGF-2) bind to the insulin-like growth factor-1 receptor (IGF-1R). The anti-IGF-1R antibody suitable for use in the present invention is preferably a monoclonal antibody, and furthermore is a chimeric antibody (human constant domain), a humanized antibody, and particularly preferably a fully human antibody. A particular anti-IGF-1R antibody suitable for use in the present invention binds to human IGF-1R' in a manner that competes with antibody 18, that is, it binds to antibody 18 to the same epitope of IGF-1R, a phenomenon described in WO 2005/005635 in. A further characteristic of the specific anti-IGF-1R antibody is that the affinity for IGF-1R is 10·8 M (KD) or less than 1〇-8 ,, especially about 1〇·9 Μ to 10-13 Μ, and preferably not A detectable concentration-dependent inhibition of insulin binding to the insulin receptor is shown. A particular anti-IGF-1R antibody suitable for use in the present invention comprises a complementarity determining region (CDR) having the sequence: a) an antibody heavy chain comprising SEQ ID NO: 144 or SEQ ID NO: 146 as CDR CDR1, CDR2 and CDR3;

b) an antibody light chain comprising SEQ ID NO: 145 or SEQ ID NO: 147 as CDR 161561.doc · 65 · 201247221 CDR1, CDR2 and CDR3. In particular, an anti-IGF-1R antibody suitable for use in the present invention comprises an antibody heavy bond variable domain amino acid sequence of SEQ ID NO: 41 and an antibody light bond variable domain amino acid sequence of SEQ ID NO: 42, or an antibody heavy The chain variable domain amino acid sequence SEQ ID NO: 43 and the antibody light chain variable domain amino acid sequence SEQ ID NO: 44. The specific anti-IGF-1R antibody suitable for use in the present invention can be obtained from a fusion tumor cell line <IGF-1R>HUMAB-pure line 18 and <IGF-1R>HUMAB-pure line 22', respectively, with the accession number DSM ACC 2587 and DSM ACC 2594 are deposited with the German Collection of Microorganisms (Deutsche Sammlung von

Mikroorganismen und Zellkulturen GmbH ; DSMZ) (Germany) 其他 Other suitable anti-IGF-1R antibodies suitable for use in the present invention are, for example, fully human IgG 1 mAb coxitumumab/IMC-A12 (described in Burtrum et al., Cancer) Res 63, 8912-21 (2003); Rowinsky et al, Clin Cancer Res 13, 5549S-5555s (2007), fully human IgGl mAb AMG-479 (described in Beltran et al, Mol Cancer Ther 8, 1095-1105) (2009); Tolcher et al, J Clin Oncol 27, 5800-7 (2009), humanized IgG1 mAb MK-0646/h7C10 (described in Goetsch et al, Int J Cancer 113, 316-28 (2005); Broussas et al, Int J Cancer 124, 2281-93 (2009); Hidalgo et al, J Clin Oncol 26, Abstract 3520 (2008); Atzori et al, J Clin Oncol 26, Abstract 3519 (2008), Humanization IgGl mAb AVE1642 (described in Descamps et al, Br J Cancer 100, 366-9 (2009); Tolcher et al, J Clin Oncol 26, Abstract 3582 (2008); Moreau et al, Blood 110, Abstract 1 166 (2007) ; Maloney et al., Cancer Res 63, 161561.doc • 66 - 201247221 5073-83 (2003)), complete person IgG2 mAb figitumumab/CP-751, 871 (Cohen et al, Clin Cancer Res 11, 2063-73 (2005); Haluska et al, Clin Cancer Res 13, 5834-40 (2007); Lacy Et al, J Clin Oncol 26, 3196-203 (2008); Gualberto and Karp, Clin Lung Cancer 10, 273-80 (2009)), fully human IgG1 mAb SCH-717454 (described in WO 2008/076257 or Kolb et al. , Pediatr Blood Cancer 50, 1190-7 (2008), 2.13.2. mAb (described in US 7,037,498 (WO 2002/053596)) or fully human IgG4 mAb BIIB022. A particular anti-CEA antibody suitable for use in the present invention is a humanized IgG class antibody having the binding specificity of a murine PR1 A3 antibody (Richman and Bodmer, Int J Cancer 3 9, 317-328 (1987)). Particularly suitable are humanized IgG class anti-CEA antibodies comprising: a) CDR1 (SEQ ID NO: 148), CDR2 (SEQ ID NO: 149) and CDR3 (SEQ ID NO: 150) in the heavy chain variable domain And b) CDR1 (SEQ ID NO: 151), CDR2 (SEQ ID NO: 152) and CDR3 (SEQ ID NO: 153) in the light chain variable domain. More specific anti-CEA antibodies suitable for use in the invention comprise the heavy chain variable domain SEQ ID NO: 154 and the light chain variable domain SEQ ID NO: 155. Such anti-CEA antibodies are described in PCT Publication No. WO 20 11/023787, the disclosure of which is incorporated herein in its entirety. Specific anti-HER3 antibodies suitable for use in the present invention are humanized IgG class antibodies, such as Mab 205.10.1, Mab 205.10.2 and Mab 205.10.3 as described in PCT Publication No. WO 2011/076683, especially Mab 161561. Doc -67- 201247221 205.10.2. A specific anti-CDCP1-antibody suitable for use in the present invention is a humanized 1 § steroid derived from (1 1-4 antibody as described in PCT Publication No. WO 2011/023 3 89 (Accession No. DSM ACC 2551 ( DSMZ)). Exemplary anti-MCSP antibodies useful in the present invention are described, for example, in WO 2006/100582. In one embodiment, the antibody is an IgG class full length antibody. In a particular embodiment, the antibody is an IgGl antibody. In an embodiment, the antibody comprises a human Fc region, more specifically a human IgG Fc region, most specifically a human IgG1 Fc region. Antibodies suitable for use in the invention, such as the anti-IGF-1R, anti-EGFR described above And an anti-CD20 antibody, which may comprise a human Igy-1 heavy chain constant region as described in SEQ ID NO: 156 (ie, the antibodies belong to the human IgG1 subclass). Antibodies suitable for use in the present invention are engineered to An engineered antibody has an increased effector function compared to an effector. In one embodiment, an antibody engineered to increase effector function is at least 2-fold, at least 10-fold, or even at least 100 in effector function compared to a corresponding unengineered antibody. Double. Effect function increased This may include, but is not limited to, one or more of the following: increased Fc receptor binding, increased Clq binding and complementation-dependent cytotoxicity (CDC), increased antibody-dependent cell-mediated cytotoxicity (ADCC), antibody dependence Increased phagocytic phagocytosis (ADCP), increased secretion of cytokines, immune complex-mediated antigens, increased uptake of antigen by cells, increased binding to NK cells, increased binding to giant sputum cells, and binding to monocytes Increased, direct binding to polymorphonuclear cells, direct signal to induce apoptosis 161561.doc .68 · 201247221 Increased conduction, increased cross-linking of antibodies bound to the target, increased dendritic cell maturation, or T cell activation (priming) In one embodiment, the effector function is increased to one or more selected from the group consisting of increased Fc receptor binding, increased CDC, increased ADCC, increased ADCP, and increased cytokine secretion. In one embodiment, effector function Increased to increase binding to an activated Fc receptor. In one such embodiment, compared to the binding affinity of the corresponding unengineered antibody, The binding affinity of the Fc receptor is increased by at least 2-fold, especially by at least 10-fold. In a particular embodiment, the activated Fc-receptor system is selected from the group consisting of FcyRIIIa, FcyRI, and FcyRIIa. In one embodiment, the activated Fc receptor is FcyRIIIa In another embodiment, the increase in effector function is an increase in ADCC. In one such embodiment, the ADCC is increased by at least 10-fold, especially at least 100-fold, compared to the ADCC mediated by the corresponding unengineered antibody. In yet another embodiment, the effector function is increased to increase binding to the activated Fc receptor and increase in ADCC. The increase in effect function can be measured by methods known in the art. Suitable assays for measuring ADCC are described herein. Further examples of in vitro assays for assessing ADCC activity of related molecules are described in U.S. Patent No. 5,500,362; Hellstrom et al, Proc Natl Acad Sci USA 83 7059-7063 (1986); and Hellstrom et al. Proc Natl Acad Sci USA 82, 1499-1502 (1985); U.S. Patent No. 5,821,337; Bruggemann et al., J Exp Med 166, 1351-1361 (1987). Alternatively, non-radioactive assays can be used (see, for example, ACTITM non-radioactive cytotoxicity analysis for flow cytometry)

161561.doc -69- S 201247221 (CellTechnology, Inc., Mountain View 'CA); and CytoTox 96® Non-Radioactive Cytotoxicity Assay (Promega ' Madison, WI)) » Applicable effector cells for these analyses include peripheral blood Monocytes (PBMC) and natural killer (NK) cells. Alternatively or additionally, the ADCC activity of the relevant molecule can be assessed in vivo, for example, in an animal model such as the animal model disclosed in Clynes et al., Proc Natl Acad Sci USA 95, 652-656 (1998). Binding to an Fc receptor can be readily determined by, for example, ELIS A or surface plasmon resonance (SPR) using standard instrument configurations such as BIAcore Instruments (GE Healthcare) and such as Fc receptors obtainable by recombinant expression. The binding affinity to an activated Fc receptor according to a specific example was measured by surface plasmon resonance using a BIACORE® T100 machine (GE Healthcare) at 25 °C. Alternatively, the binding affinity of an antibody to an Fc receptor can be assessed using a cell strain known to be used to express a particular Fc receptor, such as NK cells expressing the Fcyllla receptor. Clq binding assays can also be performed to determine if an antibody is capable of binding Clq and thus has CDC activity. See, for example, the Clq and C3c binding ELISAs in WO 2006/029879 and WO 2005/100402. To assess complement activation, CDC analysis can be performed (see, for example, Gazzano-Santoro et al, J Immunol Methods 202, 163 (1996); Cragg et al, Blood 101, 1045-1052 (2003); and Cragg and Glennie, Blood 103, 2738-2743 (2004)). An increase in effector function can be caused, for example, by glycosylation of the Fc region of the antibody or by introduction of an amino acid mutation in the Fc region of the antibody. In one embodiment, the anti-system is engineered by introducing one or more amino acid mutations in the Fc region. In a particular embodiment, the amino acid is mutated to an amino acid substitution. In a particular embodiment of 161561.doc •70·201247221 'amino acid substitutions 298, 333 and/or 334 at the Fc region (EU numbering of residues suitable for amino acid mutations are described, for example, in the literature below) : Shields et al, J Biol Chem 9 (2), 6591-6604 (2001); US Patent No. 6,737,056; WO 2004/063351 and WO 2004/099249. Mutated Fc regions can be deleted, substituted, inserted by amino acids Or modification 'prepared using genetic or chemical methods well known in the art. Genetic methods may include site-specific mutagenesis of coding DNA sequences, PCR, gene synthesis, and the like. Validation of the correct nucleus by, for example, sequencing. In other embodiments, the anti-system is engineered by modifying glycosylation in the Fc region. In a particular embodiment, the antibody is engineered to increase compared to unengineered antibodies. Proportion of non-fucosylated oligosaccharides in the Fc region. Increased proportion of non-fucosylated oligosaccharides in the Fc region of the antibody increases the effector function of the antibody, particularly increasing ADCC. In a more specific example, the antibody Fc region N in the company At least about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 500/❶, about 55%, about 60%, about 65%, about about 5%. 70%, about 75%, about 80%, about 85, about 90%, about 95% or about 100%, preferably at least about 5%, more preferably at least about 70% without trehalose. Oligosaccharides that are not hyalosylated may be of a mixed or complex type. In another specific embodiment, the antibody is engineered to increase the quaternary oligosaccharide in the Fc region compared to an unengineered antibody. In a more specific embodiment, at least about 1%, about 15%/〇, about 20〇/〇, about 25〇/〇, about 30%, about about N in the Fc region of the antibody. 35〇/〇, about 4〇%, about 16l561.doc -71· 201247221 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or about 100%, preferably at least about 50%, more preferably at least about 70% are halved. The halved oligosaccharide may be of a mixed or complex type. In the examples, the antibody is engineered to increase the Fc region compared to an unengineered antibody. The ratio of the type but not the fucosylated oligosaccharide. In a more specific embodiment, at least about 10%, about 15%, about 20%, about 25% of the N-linked oligosaccharides in the Fc region of the antibody, About 30°/., about 35°/., about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85. %, about 90%, about 95% or about 100%, preferably at least about 15%, more preferably at least about 25%, at least about 35°/. Or at least about 50% is halved without hyalcosylation. The oligosaccharides which are halved and which are not hyalcosylated may be of a mixed type or a complex type. The oligosaccharide structure in the Fc region of an antibody can be analyzed by methods well known in the art, for example by Umana et al, Nat Biotechnol 17, 176-180 (1999) or Ferrara et al, Biotechn Bioeng 93, 85. MALDI TOF mass spectrometry as described in 1-861 (2006). The percentage of non-fucosylated oligosaccharides is the amount of oligosaccharides lacking trehalose residues relative to all oligosaccharides attached to Asn 297 (eg, complex mannose structure, mixed mannose structure, and high mannose structure), and Samples treated with N-glycosidase F were identified by MALDI TOF MS. Asn 297 refers to the aspartame residue at position 297 (the EU numbering of the Fc region residues) at the Fc region; however, Asn 297 may also be located upstream or downstream of position 297 due to minor sequence variations in the antibody. ±3 amino acids, i.e. at positions 294 to 300. Determination of the percentage of oligosaccharide or split-type unfucosylated oligosaccharide in a similar manner. In one embodiment, the anti-system is altered by one or more glycosyltransferases. The antibody is produced in an active host cell and engineered to have a modified glycosylation in the Fc region compared to an unengineered antibody. Glycosyltransferases include β(1,4)-indolylglucosyltransferase ni (GnTIII), β(1,4)-galactosyltransferase (GalT), β(1,2) -Ν_ Ethyl Glucosyltransferase I (GnTI), β(1,2)-Ν-Ethyl Glucosyltransferase II (GnTII) and α(1,6)- Trehalose Transfer Enzyme. In a specific embodiment, the anti-system is engineered by the production of the antibody in a host cell with increased β(1,4)-indolylglucosyltransferase m (GnTIII) activity, thereby The proportion of unfucosylated oligosaccharides in the Fc region was increased compared to unengineered antibodies. In an even more specific embodiment, the host cell additionally has increased alpha-mannose lipase II (Manll) activity. Glycosylation engineering methods useful for engineering antibodies suitable for use in the present invention have been described in detail in Umana et al, Nat Biotechnol 17, Ρό-ΐδΟ (1999); Ferrara et al, Biotechn Bioeng 93, 851 - 861 (2006); WO 99/54342 (U.S. Patent No. 6,602,684; EP 1071700); WO 2004/065540 (U.S. Patent Application Publication No. 2004/0241817; EP 1587921), WO 03/01 1878 (US Patent Application) Publication No. 2003/01 75884, the contents of which are incorporated herein by reference in its entirety. Glycosyl engineered antibodies using this method are referred to herein as GlycoMab. In general, any type of cultured cell line, including the cell lines discussed herein, can be used to produce a cell line for producing an anti-73·16156I.doc 201247221 TNC A2 antibody having an altered glycosylation pattern. Specific cell lines include CHO cells, sputum cells, NSO cells, SP2/0 cells, sacral tumor cells, Ρ3Χ63 mouse myeloma cells, PER cells, PER.C6 cells or fusion tumor cells, and other mammalian cells. In certain embodiments, the host cell has been manipulated to exhibit an increased amount of one or more polypeptides having beta (1,4)-indolylglucosyltransferase III (GnTIII) activity. In certain embodiments, the host cell has been further manipulated to exhibit an increased amount of one or more polypeptides having alpha-mannosidase II (Manll) activity. In a specific embodiment, the polypeptide having GnTIII activity is a fusion polypeptide comprising a catalytic domain of GnTIII and a Golgi localization domain of a heterologous Golgi resident polypeptide. In particular, the Golgi localization domain is the Golgi localization domain of mannosidase II. Methods for producing such fusion polypeptides and using the same to produce antibodies with increased effector function are disclosed in Ferrara et al, Biotechn Bioeng 93, 85 pp 861 (2006) and WO2004/065540, the contents of which are expressly incorporated by reference in their entirety. The manner is incorporated herein. Host cells containing a coding sequence for an antibody of the invention and/or a coding sequence for a polypeptide having glycosyltransferase activity and which exhibit a biologically active gene product can be identified by methods well known in the art, such as DNA. -DNA hybridization or DNA-RNA hybridization; presence or absence of "marker" gene function; assessment of transcription, as measured by expression of individual mRNA transcripts in host cells; or measurement of gene products, such as by immunization Analytical methods are measured by their biological activity. The GnTIII or Man II activity can be detected, for example, by using a lectin that binds to a biosynthetic product of GnTIII or Manll, respectively. An example of such a lectin is E4-PHA lectin, which preferentially binds to an oligosaccharide containing a bipartite GlcNAc. Biosynthesis products (i.e., specific oligosaccharide structures) having a polypeptide active with GnTIII or Manll 161561.doc • 74·201247221 may also be obtained by oligosaccharides released from glycoproteins produced by cells expressing the polypeptides. Mass spectrometry to detect. Alternatively, a functional assay that measures increased effector function (e.g., increased binding of Fc receptors), which is mediated by antibodies produced by cells engineered with a polypeptide having GnTIII or Manll activity, can be used. In another embodiment, the anti-system is engineered to produce an Fc by production of the antibody in a host cell having reduced alpha(1,6)-trehalyltransferase activity, thereby increasing Fc compared to an unengineered antibody. The proportion of non-fucosylated oligosaccharides in the region. A host cell with reduced alpha(1,6)-trehalyltransferase activity may be a cell in which the alpha(1,6)-trehalyltransferase gene has been disrupted or otherwise inactivated (eg, gene knockout) (see Yamane-Ohnuki et al, Biotech Bioeng 87, 614 (2004); Kanda et al, jBz. oewg, 94 (4), 680-688 (2006); Niwa et al, J Immunol Methods 306, 151-160 (2006) Other examples of cell lines capable of producing de-fucosylated antibodies include Lecl3 CHO cells lacking protein hacosylation (Ripka et al, Arch Biochem Biophys 249, 533-545 (1986); U.S. Patent Application No. US 2003/0157108; and WO 2004/056312, especially example 11). Alternatively, antibodies suitable for use in the present invention may be in accordance with EP 1 176 195 Al, WO 03/084570, WO 03/085119, and U.S. Patent Application Publication No. 2003/0115614, No. 2004/093621, No. 2004/110282, Techniques disclosed in U.S. Patent No. 6,946,292 (Kyowa), for example, by reducing or eliminating 161561.doc-75-201247221 in addition to GDP in host cells for antibody production. - The activity of the trehalose transporter is glycosyl engineered to reduce trehalose residues in the Fc region. Glycosyl engineered antibodies suitable for use in the present invention can also be produced in expression systems that produce modified glycoproteins, such as WO 03/056914 (GlycoFi, Inc.) or WO 2004/057002 and WO 2004/024927 (Greenovation). Methods for the production of antibodies and immunoconjugates suitable for use in the present invention are well known in the art and are described, for example, in WO 2011/020783, WO 2005/044859, WO 2006/082515 ' WO 2008/017963, WO 2005/005635, WO 2008/077546 'WO 2011/023787, WO 2011/076683, WO 2011/023389 and WO 2006/100582. Determining methods for producing polyclonal antibodies and monoclonal antibodies, for example, in Harlow and Lane, "Antibodies, a laboratory manual", Cold Spring Harbor Laboratory, 1988, 0 non-naturally occurring antibodies or fragments thereof can be used in solid phase Peptide synthesis for construction, recombination (e.g., as described in U.S. Patent No. 4,816,567), or by, for example, screening a combinatorial library comprising a variable heavy chain and a variable light chain (see, for example, the McCafferty) U.S. Patent No. 5,969,108). For recombinant production of immunoconjugates and antibodies suitable for use in the invention, one or more polynucleotides encoding the immunoconjugate or antibody are isolated and inserted into one or more vectors for further selection in a host cell and/or Or performance. The polynucleotides can be readily isolated and sequenced using conventional procedures. Expression vectors containing the 161561.doc-76-201247221 coding sequence for antibodies or immunoconjugates, as well as appropriate transcription/translation control signals, can be constructed using methods well known to those skilled in the art. Such methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo recombination/genetic recombination. See, for example, Maniatis et al., MOLECULAR CLONING: A LABORATORY MANUAL, Cold Spring Harbor Laboratory, Ν·Υ· (1989) and Ausubel et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, Greene Publishing Associates and Wiley Interscience, NY (1989). Technology. Immunoconjugates suitable for use in the present invention can be represented by a single polynucleotide encoding the entire immunoconjugate or a plurality of (e.g., two or more) polynucleotides that are co-expressed. Polypeptides encoded by co-presented polynucleotides can be associated via, for example, disulfide bonds or other means to form a functional immunoconjugate. For example, the heavy chain portion of the antigen binding portion can be encoded by a polynucleotide that is separate from the portion of the immunoconjugate that comprises the light chain portion of the antigen binding portion and the portion of the effector portion. When co-presented, the heavy chain polypeptide will associate with the light chain polypeptide to form an antigen binding portion. Alternatively, in another example, the light chain portion of the antigen binding portion can be encoded by a polynucleotide that is separate from the portion of the immunoconjugate that comprises the heavy chain portion of the antigen binding portion and the portion of the effector portion. Host cells suitable for replication and support of recombinant protein expression are well known in the art. Such cells can be transfected or transduced with a specific expression vector as appropriate, and a large number of cells containing the vector can be grown for inoculation of a large-scale fermentation tank to obtain a sufficient amount of protein for use in, for example, clinical applications. Suitable host cells include prokaryotic microorganisms such as E. coli; or various eukaryotic cells such as Chinese hamster ovary cells (CHO), insect 161561.doc-77·201247221 cells or the like. For example, recombinant proteins can be produced in bacteria, especially when glycosylation is not required. After performance, the protein can be isolated from the bacterial cell paste in the soluble fraction and can be further purified. In addition to prokaryotes, eukaryotic microorganisms (such as filamentous fungi or yeast) are also suitable for the selection or expression of hosts for protein-encoding vectors, including fungal and yeast strains whose glycosylation pathway has been "humanized", resulting in partial or A fully human glycosylated version of the protein. See Gerngross, Nat Biotech 22, 1409-1414 (2004) and Li et al, Nat Biotech 24, 210-215 (2006). Host cells suitable for the expression of (glycosylated) proteins are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant cells and insect cells. A number of baculovirus strains have been identified which can be used in conjunction with insect cells, especially for transfecting Spodoptera frugiperda cells. Plant cell cultures can also be utilized as a host. See, for example, U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (the PLANTIBODIEStm technique for the production of antibodies in transgenic plants). Vertebrate cells can also be used as hosts. For example, a mammalian cell strain suitable for growth in suspension can be used. Other examples of suitable mammalian host cell strains are monkey kidney CV1 cell line (COS-7) transformed from SV40; human embryonic kidney (HEK) cell line (293 or 293T cells, eg, such as Graham et al., J Gen Viro丨) 36, 59 (1977); baby hamster kidney cells (BHK); mouse 赛toli cells (TM4 cells 'eg, as described in Mather, Biol Reprod 23, 243-251 (1980)); monkeys Renal cells (CV1); African green monkey kidney cells (VERO-76); human cervical cancer cells (HELA); canine 161561.doc -78- 201247221 kidney cells (MDCK); buffalo rat liver cells (BRL 3A) Human lung cells (W138); human hepatocytes (Hep G2); mouse mammary tumor cells (MMT 060562); TRI cells (eg, as in Mather et al., Annals NY Acad Sci 383, 44-68 (1982) Said); MRC 5 cells; and FS4 cells. Other suitable mammalian host cell lines include Chinese hamster ovary (CHO) cells, including dhfr. CHO cells (Urlaub et al, Proc Natl Acad Sci USA 77, 4216 (1980)); and myeloma cell lines, such as Y0, NS0, P3X63 and Sp2/0. For a review of certain mammalian host cell lines suitable for the production of proteins, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 255-268. (2003). Host cells include cultured cells, such as mammalian cultured cells, yeast cells, insect cells, bacterial cells, and plant cells, to name a few; and cells contained in a transgenic animal, a transgenic plant, or a cultivated plant or animal tissue. . In one embodiment, the host cell is a eukaryotic cell, particularly a mammalian cell, such as a Chinese hamster ovary (CHO) cell, a human embryonic kidney (HEK) 293 cell, or a lymphoid cell (e.g., Y0, NSO, Sp20 cells). If the antibody and immunoconjugate are intended for human use, a chimeric form of the antibody or antigen binding portion can be used, wherein the antibody constant region is from a human. The human or fully human form of the antibody or antigen-binding portion can also be prepared according to methods well known in the art (see, for example, U.S. Patent No. 5,565,332 issued toW. Humanization can be achieved by a variety of methods including, but not limited to, (a) the retention or non-retention of key framework residues (eg, residues that are important for retaining good antigen binding affinity or antibody function) 161561.doc • 79·201247221 CDRs of non-human (eg donor antibodies) are grafted into the framework and constant regions of humans (eg, receiving antibodies); (b) only non-human specificity determining regions (SDR or a-CDR; for antibodies) - residues that are critical for antigen interaction) are transplanted into the human framework and constant regions; or (c) transplant the entire non-human variable domain, but by replacing surface residues, the "cover" is intended to be part of humans. Humanized antibodies and methods for their production are reviewed, for example, in Almagro and Fransson, Front Biosci 13, 1619-1633 (2008), and are further described, for example, in Riechmann et al, Nature 332, 323-329 (1988); Et al., Proc Natl Acad Sci USA 86, 10029-10033 (1989); U.S. Patent Nos. 5,821,337, 7,527,791, 6,982,321 and 7,087,409; Jones et al, Nature 321, 522-525 (1986) Morrison et al, Proc Natl Acad Sci 81, 6851-6855 (1984); Morrison and Oi, Adv Immunol 44, 65-92 (1988); Verhoeyen et al, Science 239, 1534-1536 (1988); Padlan, Molec Immun 31 (3), 169-217 (1994); Kashmiri et al, Methods 36, 25-34 (2005) (describe SDR (a-CDR) transplantation); Padlan, Mol Immunol 28, 489-498 (1991) ( Describe "resurfacing"; Dall'Acqua et al, Methods 36, 43-60 (2005) (description "FR reorganization"); and Osbourn et al, Methods 36, 61-68 (2005); Klimka et al., Br J Cancer, 83, 252-260 (2000) (describes the "directed selection" method of FR reorganization). Human antibodies and human variable regions can be produced using a variety of techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, Curr Opin Pharmacol 5, 368-74 (2001) and Lonberg, Curr Opin Immunol 161561. doc • 80-201247221 20, 450-459 (2008). The human variable region can form part of a human monoclonal antibody produced by the fusion tumor method and can be derived from a human monoclonal antibody produced by the fusion tumor method (see, for example, Monoclonal Antibody Production Techniques and Applications, pages 51 to 63 (Marcel Dekker). , Inc., New York '1987)). Human antibodies and human variable regions can also be prepared by administering an immunogen to an engineered transgenic animal to react against an antigenic attack to produce an intact human antibody or an intact antibody having a human variable region (see, for example, Lonberg, Nat). Biotech 23, 1117-1125 (2005)). Human antibodies and human variable regions can also be produced by isolating Fv pure line variable region sequences selected from human-derived phage display libraries (see, for example, Hoogenboom et al., Methods in Molecular Biology 178, 1-37 (0' Brien et al.) Ed., Human Press, Totowa, NJ, 2001); and McCafferty et al, Nature 348, 552-554; Clackson et al, Nature 352, 624-628 (1991)). Phage typically present antibody fragments as single-chain Fv (scFv) fragments or Fab fragments. In certain embodiments, antibodies or antigen-binding portions suitable for use in the present invention are engineered to enhance binding affinity according to methods disclosed in, for example, U.S. Patent Application Publication No. 2004/0132066, the disclosure of which is incorporated herein by reference. The manner of full reference is incorporated herein. The ability of an antibody or antigen binding portion of the invention to target a particular antigenic determinant can be measured by enzyme-linked immunosorbent assay (ELISA) or other techniques well known to those skilled in the art, such as surface plasmon resonance. Techniques (analysis on the BIACORE T100 system KLiljeblad et al., Glyco J 17, 323-329 (2000)) and traditional binding analysis (Heeley, Endocr Res 28, 217-229 (2002)). 161561.doc -81 - 201247221 Antibodies and immunoconjugates prepared as described herein can be purified by techniques known in the art, such as high performance liquid chromatography, ion exchange chromatography, gel electrophoresis, affinity Chromatography, size exclusion chromatography and the like. The actual conditions used to purify a particular protein will depend, in part, on factors such as net charge, hydrophobicity, hydrophilicity, and the like, and will be readily apparent to those skilled in the art. Pharmaceutical Compositions In another aspect, the present invention provides a pharmaceutical composition comprising (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety in a pharmaceutically acceptable carrier And (b) antibodies engineered to increase effector function. These pharmaceutical compositions can be used, for example, in any of the methods of treatment described below. A pharmaceutical composition of an immunoconjugate and an antibody having increased effector function as described herein by mixing the immunoconjugate and antibody of the desired purity with one or more pharmaceutically acceptable carriers selected as appropriate ( Remington's Pharmaceutical Sciences, 18th Edition, Mack Printing Company (1990)) is prepared as a lyophilized formulation or as an aqueous solution. Pharmaceutically acceptable carriers are generally non-toxic to the recipient at the dosages and concentrations employed and include, but are not limited to, buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid And methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; gasified hexahydro quaternary ammonium; gasified benzalkonium chloride; benzethonium chloride; phenol, butanol or benzoquinone Alcohol '. p-hydroxybenzoic acid alkyl ester, such as methyl p-hydroxybenzoate or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and I6156I.doc • 82 - 201247221 m-cresol); low molecular weight (less than about 1 residue) polypeptide; protein such as serum albumin, gelatin or immunoglobulin; hydrophilic polymer such as polyvinylpyrrolidone; amino acid, Such as glycine, glutamic acid, aspartame, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrin; chelating agents, such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbus Alcohol; salt-forming counterions such as sodium; metal complexes (e.g. speech - error protein thereof); and / or non-ionic surfactant, such as polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers herein further include interstitial drug dispersing agents, such as soluble neutral active hyaluronidase glycoprotein (sHASEGP), such as human soluble PH-20 hyaluronate vinegar protein, such as rHuPH20 ( HYLENEX®, Baxter International, Inc.) Some exemplary sHASEGPs and methods of use (including rHuPH20) are described in U.S. Patent Publication Nos. 2005/0260186 and 2006/0104968. In one aspect, sHASEGP is combined with one or more other glycosaminoglycanases, such as chondroitinase. Exemplary lyophilized formulations are described in U.S. Patent No. 6,267,958. Aqueous formulations include those described in U.S. Patent Nos. 6,171,586 and WO2006/044908, and the latter formulations include histidine acetate buffer. The pharmaceutical compositions herein may also contain other active ingredients as are necessary for the particular indication being treated, especially those having complementary activities which do not adversely affect each other. For example, if the disease to be treated is cancer, it may be desirable to further provide one or more anticancer agents, such as a chemotherapeutic agent, a tumor cell proliferation inhibitor, or a tumor cell apoptosis activating agent. The 161561.doc -83 - 201247221 active ingredient is suitably present in the combination in an amount effective for the intended purpose. The active ingredient may also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polyCT methods, for example, by hydroxymethylcellulose or gelatin microcapsules and poly-(methyl methacrylate) microcapsules, respectively. In capsules, it is trapped in a gelled drug delivery system (eg, vesicles, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 18, Mack Printing Company (1990). Sustained-release preparations can be prepared. Suitable examples of sustained-release preparations include semi-permeable matrices of solid hydrophobic polymers containing antibodies in the form of shaped articles such as films or microcapsules. Compositions for administration in vivo are generally sterilized. Sterilization can be easily achieved by, for example, filtration through a sterile filtration membrane. Methods of Treatment The combination of (a) an immunoconjugate comprising at least one antigen binding portion and an effector portion and (b) an antibody engineered to increase effector function, as provided herein, can be used in a method of treatment. In one aspect, a combination comprising an immunoconjugate comprising at least one antigen binding portion and an effect portion and (b) an engineer engineered to increase effector function is provided, the combination being for use as a medicament. In another aspect, a combination of (a) an immunoconjugate comprising at least one antigen binding portion and an effector portion and an engineered antibody that increases effector function is provided for use in the treatment of a disease. In certain embodiments, a combination of (a) an antibody comprising at least one antigen binding portion and an effector, and (b) an engineered antibody that increases effector function 161561.doc-84-201247221 is provided. Used in therapeutic methods. In certain embodiments, the invention provides a combination of (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) an engineered antibody that increases effector function, the combination being used to treat a disease In an individual method, the method comprises administering the combination of therapeutic effects of the individual. In one such embodiment, the method further comprises administering to the individual a therapeutically effective amount of at least one other therapeutic agent 'e' as described below. In other embodiments, the invention provides (a) a combination of an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) an engineered antibody that increases effector function for stimulating effector cell function . In certain embodiments, the invention provides (a) a combination of an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) an engineered antibody that increases effector function, the combination being used to stimulate an individual effect In a method of cellular function, the method comprises administering to the individual an effective amount of the combination to stimulate effector cell function. An "individual" according to any of the above embodiments is a mammal, especially a human. A disease according to any of the above embodiments is a condition which can be treated by stimulating effector cell function. In certain embodiments, the disease is a cell proliferative disorder, particularly cancer. In another aspect, the invention provides (a) an immunoconjugate comprising at least one antigen-binding knife and an effect knife and (8) a modified process. The use of a combination of antibodies that increase the effect of the effect is used in the manufacture or preparation of a medicament. In one: Example: '13⁄4 drug is used to treat disease. In another embodiment, the medicament is for use in a method of treating a travel, loyalty, or sputum disease, the method comprising administering to a diseased individual a therapeutically effective amount of a sputum. _ , ', In one such embodiment, the method further comprises 161561.doc • 85· 201247221 comprising administering to the individual a therapeutically effective amount of at least one other therapeutic agent, for example as described below. In another embodiment, the medicament is for stimulating effector cell function. In another embodiment, the medicament is for use in a method of stimulating an individual's effect on cellular function. The method comprises administering to the individual an agent effective to stimulate the effector cell. The "individual body" according to any of the above embodiments is a mammal, especially a human. According to (d) the "disease" of the above embodiment is a condition which can be treated by stimulating effector cell function. In certain embodiments, the disease is a cell proliferative disorder, particularly cancer. In another aspect, the invention provides a method of treating a disease. In one embodiment, the method comprises administering to the individual having the disease a therapeutically effective amount of (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety, and an antibody that increases effector function Combinations In one such embodiment, the method further comprises administering to the individual a therapeutically effective amount of at least one other therapeutic agent as described below. An individual according to any of the above embodiments is a mammal, especially a human. A "disease" according to any of the above embodiments is a disease which can be treated by stimulating effector cell function. In certain embodiments, the disease is a cell proliferative disorder, particularly cancer. In another aspect, the invention provides methods for stimulating the function of an individual's effector cells. In one embodiment, the method comprises administering to the individual an effective amount of (a) a combination of an immunoconjugate comprising at least one antigen binding portion and an effect portion and (b) an antibody engineered to increase effector function, such that Stimulate effector cell function. In one embodiment, the "individual" is a mammal, especially a human. In another aspect, the invention provides any of the antibodies comprising (a) an immunoconjugate comprising at least one antigenic junction 161561.doc-86 - 201247221 and an effector moiety and (b) an engineered antibody that increases effector function A combined pharmaceutical composition for use in, for example, any of the above methods of treatment. In one embodiment, the pharmaceutical composition comprises (4) an immunoconjugate comprising at least one antigen binding portion and an effect portion as provided herein (bm combined with an antibody engineered to increase effector function and a pharmaceutically acceptable carrier) In another embodiment, the pharmaceutical composition comprises any combination provided herein and at least one other therapeutic agent, such as described below. According to any of the above embodiments, the disease is treatable by stimulating effector cell function The combination of the present invention is useful for treating a disease condition that is beneficial to the immune system of the host, particularly a condition that requires an enhanced cellular immune response. These may include disease conditions in which the host immune response is insufficient or absent. The combined disease condition comprises, for example, a tumor or an infection, wherein the cellular immune response will be the key to specific immunity. (4) The specific disease conditions in which the combination of the present invention can be used include cancers, particularly renal cell carcinomas or melanomas; In particular, HIV-positive patients, immunosuppressed patients, chronic infections and the like. In certain embodiments, the disease is a cell proliferative disorder in which the disease is cancer, particularly selected from the group consisting of lung cancer, colorectal cancer, renal cancer, prostate cancer, breast cancer, head and neck cancer, ovarian cancer, and brain cancer. Cancer of a group of lymphomas, leukemias, skin cancers. Combinations of the invention may be used in therapy alone or in combination with other agents. For example, 'the invention (the combination may be co-administered with at least one other therapeutic agent. In some embodiments, 'other therapeutic agents are anti-cancer! For example, chemotherapeutic agents' tumor cell proliferation inhibitors; or tumor cells> weekly death activators. 161561.doc -87 · 201247221 Together with antibodies and immunological binding

Administration intradermally, intraperitoneally or subcutaneously. Combination therapies as provided herein encompass the administration of a subject together (where two or more therapeutic agents are included in the same formulation) and separate administration of antibodies and immunoconjugates (infusion including intramuscular, intravenous, arterial) The antibody and the immunoconjugate may be administered by the same or different routes, and may be administered by any appropriate route (for example, by injection, such as intravenous or subcutaneous injection), and the administration may be short-term administration or long-term administration. A variety of dosing schedules are contemplated, including but not limited to single administration or multiple administrations at different time points, bolus administration, and pulse infusion. Formulation, administration, and administration of a combination of the invention in a manner consistent with good medical practice. Factors considered in this context include the particular condition being treated, the particular mammal being treated, the clinical condition of the individual patient, the etiology of the condition, the agent delivery site, the method of administration, the time course of administration, and the medical practitioner's known Other factors. The antibody is not required but may optionally be formulated with one or more agents currently used to prevent or treat the condition. The effective amount of the agent will depend on the amount of antibody and immunoconjugate present in the formulation, the type of disorder or treatment, and other factors discussed above. Such agents are typically administered at the same dosages as described herein and utilized as The routes of administration described herein are used or used at about 1% to 99% of the dosages described herein, or in any dosage and by any route that is empirically/clinically determined to be appropriate. 161561.doc -88 · 201247221 For prevention Or treating a disease, an appropriate dose of the antibody and immunoconjugate (when used in combination with the invention, optionally with one or more additional therapeutic agents) will depend on the type of disease, antibody, and type of immunoconjugate to be treated , the severity of the disease and the course of the disease, the combination of which is used for prophylactic or therapeutic purposes, prior therapy, clinical history of the patient and response to antibodies and/or immunoconjugates, and judgment of the attending physician. Antibody and immune binding Suitable for one-time or appropriate treatment of patients through a series of treatments. Depending on the type and severity of the disease, the initial candidate dose for the patient is administered. Uot; T is from about 1 pg/kg to 15 mg/kg (eg, 〇·ι mg/kg to 1 〇mg/kg) of the antibody, either by separate administration, for example by one or more times, or by continuous infusion. Dosing. A typical sputum dose can range from about !pg/kg to 1〇〇mg/kpl1〇〇mg/kg, depending on the factors mentioned above. For days or longer Repeated administration, depending on the condition, treatment will generally continue until the disease symptoms are inhibited. An exemplary antibody dose will range from about 0.05 mg/kg to about 1 mg/kg. One or more doses (or any combination thereof) of 5 mg/kg, 2.0 mg/kg, 4.0 mg/kg or 1 mg/kg 2 are administered to the patient. The doses may be administered intermittently, for example, weekly or per Three weeks (eg, to allow the patient to receive about 2 to about 2 or, for example, about 6 doses of the antibody). A higher starting dose can be administered initially, followed by one or more lower doses. An exemplary dosing regimen comprises administering an initial starting dose of the antibody at about 4 mg/kg followed by administration of a weekly maintenance dose of about 2 mg/kg of the antibody. The same considerations regarding dosage are applied to the immunoconjugates used in the combinations of the invention. However, other dosing regimens can be used. The process of this therapy is monitored by conventional techniques and analytical methods. 161561.doc -89 - 201247221 Articles of Manufacture In another aspect of the invention, an article of manufacture containing a substance useful for treating, preventing, and/or diagnosing the above conditions is provided. The article comprises one or more containers and a label or package insert on or accompanying the container: suitable containers include, for example, bottles, vials, syringes, 1 solution bags, and the like. The device can be formed from various materials such as glass or plastic. Container containment group: a composition that itself or in combination with another composition effective for treating, preventing, and/or diagnosing a condition, and may have a sterile access port (eg, the container may have a needle that can be pierced by a hypodermic needle) Intravenous solution bag or vial of the stopper). At least one active agent in the composition is an anti-H active agent used in the combination of the invention, which is an immunoconjugate used in the combination of the invention, the pot may be in the same composition and container as the antibody' or may be provided in different groups: And in the container. The label or package insert indicates that the composition is used to treat the condition. h In one aspect, the invention provides a kit intended for use in the treatment of a disease, the kit comprising (4) an immunoconjugate comprising at least one antigen binding portion and an effect portion, and (8) An antibody that is engineered to increase effector function, and optionally includes (4) package inserts. The package insert contains printed instructions for directing the use of the combination therapy as a method of treating disease. Further, the kit may comprise (4) a first container containing a composition thereof, wherein the composition comprises an antibody that is modified by a regimen to increase effector function; (b) a second container comprising the composition, wherein the composition An immunoconjugate comprising at least one antigen binding moiety and an effector moiety; and optionally, (4) a third container comprising the composition thereof. 161561.doc 201247221 The composition comprises other cytotoxic agents or other therapeutic agents. The kit of this embodiment of the invention may further comprise a package insert indicating that the composition is useful for treating a particular condition. Alternatively or additionally, the kit may further comprise a third (or fourth) container 'containing a pharmaceutically acceptable buffer' such as bacteriostatic water for injection (BWFI), phosphate buffered saline, forest Ringer's solution and dextrose solution. It may further include other materials required from a commercial and user perspective, including other buffers, diluents, filters, needles, and syringes. EXAMPLES The following are examples of the methods and compositions of the present invention. It is understood that various other embodiments may be implemented in the light of the general description provided. General Methods Glycosylation engineering of the Fc region of the antibody results in increased binding affinity to human F (RIII receptor), increased binding affinity to human FcyRIII receptor and conversion to ADCC induction and enhanced anti-tumor efficacy, and human FcyRIH receptors are expressed in Macrophages, neutrophils and natural killer (NK) cells, dendritic cells, and γδ T cells. The most widely used preclinical efficacy test substance in mice, the mouse FcyRIV (ie, the mouse of human FcyRIIIa) The homologues are present on macrosatellite cells and neutrophils instead of on NK cells. Therefore, the maximum extent of any expected improvement in the efficacy of glycosyl engineered antibodies is not reflected in these models. Human F (nrRiiia (CD16a) transgenic mouse, which exhibits stable expression of human cdi6a on murine NK cells in blood, lymphoid tissues and tumors. Furthermore, these transgenic mice are not exposed to blood. The amount of human CD16a on stimulated NK cells is reversed. 161561.doc •91 · 201247221 The amount of expression seen in humans. We also show that tumor-associated NK after antibody therapy Down-regulation of human FcyRIIIa is associated with anti-tumor activity. Finally, we demonstrated that this new mouse strain significantly improved glycosyl-engineered antibody therapy in tumors compared to human CD16-negative littermates (littermate). Efficacy in the model. Example 1 A549 lung xenograft model 2B10 targeting TNC A2 was tested in human non-small cell lung cancer (NSCLC) cell line A549 injected intravenously into SCID-human FcYRIII (hCD16) transgenic mice. Fab-IL-2-Fab immunoconjugates (SEQ ID No 117 and SEQ ID No 120) and anti-EGFR GlycoMab (SEQ ID No 142 and SEQ ID No 143). This tumor model was shown to the tendon by IHC against fresh frozen tissue. The A2 domain of protein C is positive. A549 NSCLC cells were originally obtained from ATCC (CCL-185) and deposited in the Glycart internal cell bank after amplification. Usually at 37 ° C in water saturated atmosphere / 5% In C02, tumor cell lines were cultured in DMEM containing 10% FCS (Gibco). Transplantation was performed using passage 8 with a viability of 98%. 5x106 cells/animal in 200 μΐ Aim V cell culture medium (Gibco) Intravenous injection into the tail vein. According to the promised guidelines (GV-Solas; Felasa; TierschG), female SCID-FcyRIII mice (GLYCART-RCC) (in RCC (Switzerland) breeding) with 8 to 9 weeks of age at the start of the experiment were maintained for 12 hours. Light/12 hours of dark daily circulation without specific pathogen conditions. The experimental research protocol was reviewed and approved by the local government (P 2008016). After the animals arrive, they are kept for one week to get used to the new environment and observe. Regular continuous health surveillance 161561.doc -92· 201247221 test. On study day 0, mice were injected intravenously with 5 χ丨〇 6 A549 cells, randomized and weighed. One week after tumor cell injection, mice were injected intravenously

2B10 Fab-IL-2-Fab immunoconjugate was administered twice a week for 3 weeks, anti-EGFR GlycoMab was administered once a week for 3 weeks, or 2B1〇FabIL2Fab immunoconjugate was administered twice weekly for 3 weeks with anti-EGFR Glyc〇Mab A combination of 3 weeks per week. All mice were injected intravenously with 2 dips and appropriate solutions. The dose is illustrated in Table 2. Mice in the vehicle group were injected with pBS, and mice in the treated group were injected with 2B10 Fab-IL-2-Fab immunoconjugate or anti-EGFR

GlycoMab or 2B10 Fab-IL-2-Fab immunoconjugates and anti-EGFR

A combination of GlycoMab. To obtain an appropriate amount of immunoconjugate per 2 μ μ1, the stock solution was diluted with PBS if necessary. Figure 1 shows 2B1〇 Fab_IL 2_

The combination of Fab immunoconjugates and anti-EGFR-GlycoMab mediates superior efficacy in hCD16 transgenic SCID mice, resulting in median survival and overall survival compared to 2B10 Fab-IL-2-Fab immunoconjugate or anti-egfr alone

GlycoMab is synergistically increased. Table 2 Compound dose/mouse formulation buffer IS (mg/mL) Anti-EGFR Glycomab 625 pg 20 mM His/HisCl 240 mM trehalose 0.02% Tween 20 pH 6.0 9.7 (=stock solution 丨huTNC A2 2B10 (G65S) Fab-IL2- Fab=2B10 16 pg 25 mM acid clock 125 mM NaCl, 100 mM glycine, pH 6.7 1.86 —— (= stock solution 丨---- Example 2 LS174T colorectal xenograft model 161561.doc •93· 201247221 in the spleen The 2B10 Fab-IL-2-Fab immunoconjugate targeting TNC A2 and the anti-EGFR GlycoMab were tested in a human colorectal LS174T cell line injected into SCID mice. This tumor model was shown to the tendon protein by IHC against fresh frozen tissue. The A2 domain of C is positive. LS174T cells (human colon cancer cells) were originally obtained from ECACC (European Collection of Cell Culture) and deposited in the Glycart internal cell bank after amplification. LS174T was cultured in MEM Eagle's medium (Eagle, s medium) containing 1〇0/.fcs (PAA Laboratories, Austria), 1% Glutamax, and 1% MEM non-essential amino acid (Sigma). Water saturated atmosphere, 5% c〇2 The cells were cultured, and the intramuscular injection was performed in the 18th passage in vitro with a viability of 97%. A small opening was made in the left abdomen of the anesthetized SCID mice, and 5 μL of the cell suspension was injected directly below the spleen sac via the abdominal wall ( 3><1〇6^^174 cells in the 811^ medium.) Use a clamp to close the skin wound. According to the promised guidelines (GV_s〇las; Felasa; TierschG), the experiment starts at 8 Female scm mice up to 9 weeks old (purchased from Taconics, Denmark) were maintained under specific pathogen-free conditions with a daily cycle of 12 hours light/12 hours dark. The experimental study protocol was reviewed and approved by the local government (p 2 〇〇8〇16). After the animal arrives, it is maintained for one week to get used to the new environment and observed. Continuous health monitoring is carried out regularly. On the third day of the study, the mice were intraperitoneally injected with 3χ1〇6 lsi74t cells, randomized into groups. Weighing. One week after tumor cell injection, the mice were injected intravenously with 2B10 Fab-IL-2-Fab immunoconjugate twice a week for 3 weeks, anti-EGFR Glyc〇Mab once a week for 3 weeks, or 2 m〇 FabiL2-Fab immunological binding Twice a week for 3 weeks with anti-egfr Giyc〇Mab weekly 161561.doc •94· 201247221 A combination of 3 weeks. All mice were injected intravenously with 200 μΐ of the appropriate solution. The dose is illustrated in Table 3. Mice in the vehicle group were injected with PBS, and mice in the treatment group were injected with 2Β10 Fab-IL-2-Fab immunoconjugate or anti-EGFR GlycoMab or 2Β10 Fab-IL-2-Fab immunoconjugate with anti-EGFR GlycoMab combination. To obtain an appropriate amount of immunoconjugate per 200 μM, the stock solution was diluted with PBS if necessary. Figure 2 shows the combination of 2Β10 Fab-IL-2-Fab immunoconjugates with anti-EGFR GlycoMab and 2Β10 Fab-IL-2-Fab immunoconjugate or anti-EGFR GlycoMab in terms of increased median survival and overall survival. Compared to mediating excellent efficacy. Table 3 Compound dose/mouse formulation buffer concentration (rng/mL) Anti-EGFR Glycomab 625 μΕ 20 mM His/HisCl 240 mM trehalose 0.02% Tween 20 pH 6.0 9.7 (=stock solution) huTNC A2 2B10 (G65S) Fab- IL-2-Fab = 2B10 16 gg 25 mM acid slant 125 mM NaCl, 100 mM glycine, pH 6.7 1.86 (= stock solution) Example 3 ACHN renal cancer xenograft model in human kidney injected into the SCID mouse by kidney The 3F2 Fab-IL-2-Fab immunoconjugate (SEQ ID No 102 and SEQ ID No 112) targeting the FAP and the anti-EGFR GlycoMab were tested in the cell line ACHN. This tumor model was shown to be positive for FAP by IHC against fresh frozen tissue. ACHN cells (human renal adenocarcinoma cells) were originally obtained from ATCC (American Type Culture Collection) and deposited in the Glycart internal 161561.doc-95-201247221 cell bank after amplification. The ACHN cells were cultured in DMEM containing 10% FCS in a water-saturated atmosphere/5% CO 2 at 37 ° C. The intrauterine injection was performed using the 9th passage in vitro with a viability of 97.7%. A small opening (2 cm) was cut in the right ventral and peritoneal wall of anesthetized SCID mice. 50 μM of cell suspension (1 x 16 ACH 6 ACHN cells in AimV medium) was injected into the kidney 2 mm below the capsule. Use a clamp to close the skin wound and the peritoneal wall. According to the promised guidelines (GV-Solas; Felasa; TierschG), female SCID mice (purchased from Charles River, Sulzfeld, Germany) aged 8 to 9 weeks from the start of the experiment were maintained with 12 hours light/12 hours darkness. The daily cycle is free of specific pathogen conditions. The experimental research protocol was reviewed and approved by the local government (P 2008016). After the animals arrive, they are kept for one week to get used to the new environment and observe. Continuous health monitoring is performed on a regular basis. On study day 0, mice were injected intrarenally with 1 X 106 ACHN cells, randomized and weighed. One week after tumor cell injection, mice were injected intravenously with 3F2 Fab-IL-2-Fab immunoconjugate twice a week for 3 weeks, anti-EGFR GlycoMab once a week for 3 weeks, or 3F2 Fab-IL-2-Fab The immunoconjugate was combined twice a week for 3 weeks with anti-EGFR GlycoMab once a week for 3 weeks. All mice were injected intravenously with 200 μM appropriate solution. The doses in the vehicle group were injected into PBS, and the mice in the treated group were injected with 3F2 Fab-IL-2-Fab immunoconjugate, anti-antibody. Combination of an EGFR GlycoMab or 3F2 Fab-IL-2-Fab immunoconjugate with an anti-EGFR GlycoMab. To obtain an appropriate amount of immunoconjugate per 200 μM, the stock solution was diluted with PBS if necessary. Figure 3 shows that the combination of 3F2 Fab-IL-2-Fab immunoconjugate and anti-EGFR GlycoMab causes median survival and overall survival in SCID mice with 161561.doc •96· 201247221 3F2 Fab-IL-2- Fab immunoconjugates are synergistically increased compared to anti-EGFR GlycoMab. Table 4 Compound Dose/Mouse Blending Buffer Concentration (mg/mL) Anti-EGFR Glycomab 625 \ig 20 mM His/HisCl 240 mM Trehalose 0.02% Tween 20 pH 6.0 9.7 (=Reservoir Solution) FAP 3F2 Fab-IL-2 -Fab =FAP 3F2 16 pg 25 mM acid-loading 125 mM NaCl, 100 mM glycine, pH 6.7 2.46 (=stock solution) Example 4 ACHN renal cancer xenograft model infused with SCID-human FcyRIII transgenic mice The 3F2 Fab-IL-2-Fab immunoconjugate targeting FAP and the anti-EGFR GlycoMab were tested in the human kidney cell strain ACHN. This tumor model was positive for FAP by IHC against fresh frozen tissue. ACHN cells (human renal adenocarcinoma cells) were originally obtained from ATCC (American Type Culture Collection) and deposited in the Glycart internal cell bank after amplification. ACHN cells were cultured in DMEM containing 10% FCS in water saturated atmosphere / 5% CO 2 at 37 °C. Intra-renal injection was performed using the 11th generation in vitro with a viability of 96.7%. A small opening (2 cm) was cut in the right ventral and peritoneal wall of anesthetized SCID mice. A 50 μM cell suspension (1 x 10 6 ACHN cells in AimV medium) was injected into the kidney 2 mm below the capsule. Use a clamp to close the skin wound and the peritoneal wall. According to the promised guidelines (GV-Solas; Felasa; TierschG), female SCID-FcYIII mice (GLYCART-RCC) at the beginning of the experiment were 8 to 9 weeks old 161561.doc •97· 201247221 (in RCC (Switzerland) Breeding) was maintained under specific pathogen-free conditions with a daily cycle of 12 hours light/12 hours dark. The experimental research protocol was reviewed and approved by the local government (P 2008016). Once the animal arrives, keep it for a week to get used to the new environment and observe. Continuous health monitoring is performed on a regular basis. On study day 0, mice were injected intrarenally with lxl〇6 ACHN cells, randomized and weighed. One week after tumor cell injection, mice were injected intravenously with 3F2 Fab-IL-2-Fab immunoconjugate twice a week for 3 weeks, anti-EGFR GlycoMab once a week for 3 weeks, or 3F2 Fab-IL-2-Fab The immunoconjugate was combined twice a week for 3 weeks with anti-EGFR GlycoMab once a week for 3 weeks. All mice were injected intravenously with 200 μΐ of the appropriate solution. The dose is indicated in Table 5. Mice in the vehicle group were injected with PBS, and mice in the treatment group were injected with 3F2 Fab-IL-2-Fab immunoconjugate, anti-EGFR GlycoMab or 3F2 Fab-IL-2-Fab immunoconjugate and anti-EGFR GlycoMab combination. To obtain an appropriate amount of immunoconjugate per 200 μM, the stock solution was diluted with PBS if necessary. Figure 4 shows that the combination of 3F2 Fab-IL-2-Fab immunoconjugates with anti-EGFR GlycoMab mediates superior efficacy compared to 3F2 Fab-IL-2-Fab immunoconjugate or anti-EGFR GlycoMab in terms of overall survival. . Table 5 Compound dose/mouse formulation buffer concentration (mg/mL) Anti-EGFR Glycomab 625 pg 20 mM His/HisCl 240 mM trehalose 0.02% Tween 20 pH 6.0 9.7 (=stock solution) FAP 3F2 Fab-IL-2- Fab = FAP 3F2 16 pg 25 mM dish acid if 125 mM NaCl, 100 mM glycine, pH 6.7 2.46 (= stock solution) 161561.doc 98 - 201247221 Example 5 Z138 mantle cell lymphoma xenograft model infusion of SCID intravenously - 2B10 targeting TNC A2 in human mantle cell lymphoma cell line Z138 in human FcyRIII transgenic mice

Fab-IL-2-Fab immunoconjugate and anti-CD20 GlycoMab (SEQ ID No 134 and SEQ ID No 135). This tumor model was shown to be positive for TNC A2 by IHC against fresh frozen tissue. The Z138 human mantle cell lymphoma cells were originally obtained from Professor Martin Dyer (MRC Toxicology Unit, Leicester, UK) and deposited in the Glycart internal cell bank after amplification. Tumor cell lines were usually cultured in DMEM containing 1% FCS (Gibco) in water saturated atmosphere / 50 / 〇 C02 at 37 °C. Using the 18th generation for transplantation, the viability was 98%. The l〇xl〇6 cells/animal in 200 μΐ Aim V cell culture medium (Gibco) was intravenously injected into the tail vein. According to the promised guidelines (GV-Solas; Felasa; TierschG), female SCID-Fcylll mice (GLYCART-RCC) (in RCC (Switzerland) breeding) with 8 to 9 weeks of age at the start of the experiment were maintained for 12 hours. Light/12 hours of dark daily circulation without specific pathogen conditions. The experimental research protocol was reviewed and approved by the local government (P 2008016). After the animals arrive, they are maintained for one week to get used to the new environment and observed, and regular continuous health monitoring. On day 0 of the study, mice were intravenously injected with 10 < 106 2138 cells, randomly grouped and weighed. One week after tumor cell injection, mice were injected intravenously with 2B10 Fab-lL-2-Fab immunoconjugate twice a week for 3 weeks against cd2〇.

GlycoMab is administered once a week for 3 weeks, or 2B1〇 Fab_IL 2 Fab immunoconjugates are administered twice a week for 3 weeks with anti-CD20 GlycoMab once a week for 3 161561.doc •99·201247221 weeks. All mice were injected intravenously with 200 μΐ of the appropriate solution. The dosage is illustrated in Table 6. Mice in the vehicle group were injected with PBS, and mice in the treated group were injected with 2Β10 Fab-IL-2-Fab immunoconjugate, anti-CD20 GlycoMab, or 2Β10 Fab-IL-2-Fab immunoconjugate with anti-CD20 GlycoMab The combination. To obtain an appropriate amount of immunoconjugate per 200 μM, the stock solution was diluted with PBS if necessary. Figure 5 shows the combination of 2Β10 Fab-IL-2-Fab immunoconjugates with anti-CD20 GlycoMab and 2Β10 Fab-IL-2-Fab immunoconjugate or anti-CD20 GlycoMab phase in terms of median survival and overall survival. Better than synergistic enhancement. Table 6 Compound Dose/Mouse Blending Buffer Concentration (mg/mL) Anti-CD20 Glycomab 625 pg 20 mM His/HisCl 140 mM NaCl 0.02% Tween 20 pH 6.0 10.50 (=stock solution) huTNC A2 2B10 (G65S) Fab-IL2-Fab = 2B10 16 pg 25 mM potassium hydride 125 mM NaCl, 100 mM glycine, pH 6.7 1.86 (= stock solution) Example 6 ACHN renal cancer xenograft model in humans injected with SCID-human FcyRIII transgenic mice The 28H1 Fab-IL2-Fab immunoconjugate targeting FAP and the anti-EGFR GlycoMab were tested in the renal cell line ACHN, and the immunoconjugate comprises an IL-2 tetraploid mutant (qm) lacking binding to CD25 (SEQ ID NO: 108, wherein the IL-2 sequence (SEQ ID NO: 1) is substituted with SEQ ID NO: 2; and SEQ ID NO: 113). This tumor model was positive for 161561.doc -100· 201247221 FAP by IHC against fresh frozen tissue. ACHN cells (human renal adenocarcinoma cells) were originally obtained from ATCC (American Type Culture Collection) and deposited in the Glycart internal cell bank after amplification. ACHN was cultured in DMEM containing 10% FCS in a water-saturated atmosphere / 5% CO 2 at 37 °C. Intrarenal injection was performed using the 18th generation in vitro with a viability of 97%. A small opening (2 cm) was cut in the right ventral and peritoneal wall of anesthetized SCID mice. A 50 μM cell suspension (1 x 10 6 ACHN cells in AimV medium) was injected into the kidney 2 mm below the capsule. Use a clamp to close the skin wound and the peritoneal wall. According to the promised guidelines (GV-Solas; Felasa; TierschG), female SCID-FcYIII mice (GLYCART-RCC) (in RCC (Switzerland) breeding) with 8 to 9 weeks of age at the start of the experiment were maintained for 12 hours. Light/12 hours of dark daily circulation without specific pathogen conditions. The experimental research protocol was reviewed and approved by the local government (P 2008016). After the animals arrive, they are kept for one week to get used to the new environment and observe. Continuous health monitoring is performed on a regular basis. On study day 0, mice were injected intrarenally into 6 ACHN cells, randomized and weighed. One week after tumor cell injection, mice were injected intravenously with 28H1 Fab-IL-2 qm-Fab immunoconjugate three times a week for 3 weeks, anti-EGFR GlycoMab once a week for 3 weeks, or 28H1 Fab-IL-2 qm- Fab immunoconjugates were combined three times a week for 3 weeks with anti-EGFR GlycoMab once a week for 3 weeks. All mice were injected intravenously with 200 μM of the appropriate solution. The doses are indicated in Table 7. Mice in the vehicle group were injected with PBS, and mice in the treatment group were injected with 28Η1 Fab-IL-2 qm-Fab immunoconjugate, anti-EGFR GlycoMab, or 28Η1 Fab-IL-2 qm-Fab immunoconjugate and anti-conjugate A combination of EGFR GlycoMab. To obtain an appropriate amount of immunoconjugate per 200 μM, dilute 161561.doc -101 - 201247221 stock solution with PBS if necessary. Figure 6 shows that the combination of the 28H1 Fab-IL-2 qm-Fab immunoconjugate with anti-EGFR GlycoMab is mediated compared to the 28H1 Fab-IL_2 qm-Fab immunoconjugate or anti-EGFR GlycoMab alone in terms of increased median survival. Excellent effect. Table 7 Compound Dose/Mouse Blending Buffer Concentration (mg/mL) Anti-EGFR Glycomab 625 pg 20 mM His/HisCl" 240 mM Trehalose 0.02% Tween 20 pH 6.0 9.7 (=stock solution) FAP 28H1 Fab-IL2 qm- Fab 30 pg 25 mM acid-loading 125 mM NaCl, 100 mM glycine, pH 6.7 2.74 (= stock solution) Example 7 In vitro enhanced NK cell killing ability and NK cell IFN-γ released by IL-2 immunoconjugate To determine the effect of immunoconjugates on NK cells, we assessed the kill rate of tumor cells and the release of IFN-γ by NK cells after treatment with immunoconjugates, particularly immunoconjugates containing IL-2 as an effector. For this purpose, peripheral blood mononuclear cells (PBMC) were isolated using Histopaque-1077 (Sigma Diagnostics Inc., St. Louis, MO, USA) according to standard procedures. Briefly, venous blood is obtained from healthy volunteers using a heparinized syringe. The blood was diluted 2:1 with PBS without calcium or magnesium and layered on Histopaque-1077. The gradient was centrifuged for 30 minutes at room temperature (RT) at 45 Torr xg. The boundary phase containing PBMC was collected and washed with PBS for a total of 3 times (35 〇 x g, followed by 30 〇 x g, 10 minutes, room temperature). 161S61.doc -102- 201247221 In the first experiment, isolated PBMCs were cultured together with different concentrations of IL-2 (Proleukin) or IL-2 immunoconjugates (including wild-type or quadruple mutants (qm) IL-2 targeting 28H1 Fab-IL2-Fab of FAP). Two experimental settings were tested: "in solution" in which the IL-2-containing construct was added to the cell supernatant; and "coating" in which the IL-2-containing construct was bound to the FAP, and the FAP was pre-coated. On a 96F well plate (500 Ng/well in PBS, 20 hours, 4 °C). The unbound immunoconjugate was removed by washing, followed by the addition of PBMC. In both cases, PBMC were pretreated with IL-2 containing constructs for 48 hours, then recovered and used to kill K562 stem cells with a 10:1 effector:target cell ratio (E:T). 4 hours. The killer rate of the stem cells (Roche cytotoxicity detection kit LDH) was detected by measuring the LDH released into the cell supernatant. Figure 7 shows that K562 tumor cell killing rate increases after treatment of effector cells (PBMC) in solution (A) or IL-2 constructs coated in cell plate (B) compared to untreated PBMC. . IL-2 and Fab-IL2-Fab immunoconjugates enhance the ability of PBMCs to kill target cells. In the second experiment, the isolated PBMCs were incubated with IL-2 (prolactin) or IL-2 immunoconjugates and added to the cell supernatant for 45 hours. Subsequently, PBMC were recovered and used for 10:1 E:T for anti-EGFR GlycoMab-mediated ADCC of A549 cells for 4 hours. The killer rate of the target cells (Roche cytotoxicity detection kit LDH) was detected by measuring the LDH released into the cell supernatant. Figure 8 shows 28H1 Fab-IL2-Fab pre-treated or not treated with 57 nM of targeted FAP containing wild-type (wt) or quadruple mutant (qm) IL-2 in the presence of different concentrations of anti-EGFR GlycoMab Overall A549 tumor cell kill rate due to pretreated PBMC. The results showed that almost 100% of the stem cell killing rate was obtained with the combination of the immunoconjugate and GlycoMab, and this result could not be achieved by any single agent under the current experimental conditions. Two immunoconjugates comprising wild-type or quadruple mutant IL-2 are equally effective. In another experiment, isolated PBMCs with two different inferior (5 ng/ml and 500 ng/ml) anti-EGFR GlycoMab and no sugar were used in ACR cells with 5:1 E:T in ADCC analysis. The engineered anti-EGFR antibody (Erbitux) was maintained for 21 hours. At the end of the incubation period, the iFN-γ ELISA kit (BD #550612) was used to detect IFN-γ released by PBMC into the cell supernatant. Figure 9 shows that no significant IFN-γ release was detected after incubation with antibodies alone, but in the presence of IL-2 (prolactin), 28H1 Fab-IL2-Fab or 28H1 Fab-IL2 qm-Fab, (A) Anti-EGFR GlycoMab-mediated ADCC and (B) Erbitux-mediated ADCC strongly enhance IFN-γ release during incubation time. In summary, and especially at lower antibody concentrations (5 ng/ml) and highest IL-2 (immunoconjugate) concentrations (1140 nM), IFN-γ release from anti-EGFR GlycoMab was higher than Erbitux. Finally, IFN-γ release from PBMC was determined following incubation with IL-2 (prolactin), 28H1 Fab-IL2-Fab or 28H1 Fab_IL2 qm-Fab (but without any antibodies). The experimental conditions are as described above. As shown in Figure 1 ’ IL-2 (immunoconjugate) also enhanced IFN-γ release from PBMC in the absence of ADCC-inducible antibodies. The IFN-γ content was similar to that measured in the presence of 5 ng/ml Erbitux (see Figure 9B) but below the presence of anti-EGFR GlycoMab (see Figure 9A). 161561.doc • 104· 201247221 The present invention has been described in detail above by way of illustration and example, and the description and examples should not be construed as limiting the scope of the invention. The disclosures of all patents and scientific literature cited herein are hereby expressly incorporated by reference in their entirety. [Simplified Schematic] Figure 1. 2B10 Fab-IL-2- targeting TNC A2 in human non-small cell lung cancer (NSCLC) cell line A549 injected intravenously with SCID-human FcyRIII transgenic mice Fab immunoconjugate and anti-EGFR GlycoMab. This tumor model was positive for the A2 domain of tendon protein C by IHC against fresh frozen tissue. The data show that the combination of 2B10 Fab-IL-2-Fab immunoconjugate and anti-EGFR GlycoMab is compared to 2B 10 Fab-IL-2-Fab immunoconjugate or anti-EGFR GlycoMab in terms of increasing median survival. Good efficacy (see Example 1). Figure 2. Detection of 2B10 Fab-IL_2-Fab immunoconjugates targeting TNC A2 and anti-EGFR GlycoMab in human colorectal LSI 74T cell lines injected into SCID mice via spleen. This tumor model was positive for the A2 domain of tendon protein C by IHC against fresh cold tissue. The data show that the combination of 2B10 Fab-IL-2-Fab immunoconjugates with anti-EGFR GlycoMab and 2B10 Fab-IL-2-Fab immunoconjugate or anti-EGFR GlycoMab alone in terms of increased median survival and overall survival Compared to mediating excellent efficacy (see Example 2) ^

Figure 3. 3F2 Fab-IL-2-Fab immunoconjugates targeting anti-FAP and anti-EGFR GlycoMab were tested in human kidney cell line ACHN injected intrarenally into SCID mice. IHC of fresh frozen tissue showed that this tumor model was positive for FAP 161561.doc -105· 201247221. The data show that the combination of 3F2 Fab-IL-2-Fab immunoconjugate and anti-EGFR GlycoMab synergizes with 3F2 Fab-IL-2-Fab immunoconjugate or anti-EGFR GlycoMab alone to cause median survival of SCID mice and Overall survival increased (see Example 3). Figure 4. 3F2 Fab-IL-2-Fab immunoconjugates targeting anti-FAP and anti-EGFR GlycoMab were tested in human kidney cell line ACHN injected intrarenally into SCID-human FcyRIII transgenic mice. This tumor model was positive for FAP by IHC against fresh cold bundle tissue. The data show that the combination of '3F2 Fab-IL-2-Fab immunoconjugates and anti-EGFR GlycoMab mediates superior efficacy compared to 3F2 Fab-IL-2-Fab immunoconjugate alone or anti-EGFR GlycoMab in terms of overall survival. (See Example 4). Figure 5. Detection of 2B10 Fab-IL-2-Fab immunoconjugates targeting TNC A2 and anti-CD20 GlycoMab in human mantle cell lymphoma cell line Z13 8 injected intravenously into SCID-human FcyRIII transgenic mice . This tumor model was shown to be positive for TNC A2 by IHC against fresh frozen tissue. The data show that the combination of 2B10 Fab-IL-2-Fab immunoconjugates with anti-CD20-GlycoMab synergistically increases median survival and overallity compared to 2B 10 Fab-IL-2-Fab immunoconjugate alone or anti-CD20-GlycoMab Survival rate (see Example 5). Figure 6. Detection of 28H1 Fab-IL-2-Fab immunoconjugates targeting FAP in a human kidney cell line ACHN injected intrarenally into SCID-human FcyRIII transgenic mice (containing IL lacking binding to CD25) -2 quadruple mutant (qm)) and anti-EGFR GlycoMab. This tumor model was positive for FAP by IHC against fresh frozen tissue. The data show that in terms of increased median survival 161561.doc •106-201247221, the combination of 28H1 Fab-IL-2 qm-Fab immunoconjugate and anti-EGFR GlycoMab is immunoconjugated with 28H1 Fab-IL-2 qm-Fab alone. The agent or EGFR GlycoMab mediates superior efficacy (see Example 6). Figure 7. PBMC pretreated with IL-2 (prolactin), 28H1 Fab-IL2-Fab or 28H1 Fab-IL2 qm-Fab present in solution (A) or in cell disk (B) for 48 hours. (E: T = 10: 1, 4 hours) increased K562 tumor cell kill rate. Values indicate an increase in kill rate (%) compared to untreated PBMC (see Example 8). Figure 8. PBMC pretreated with or without pretreatment of 57H1 Fab-IL2-Fab or 28H1 Fab-IL2 qm-Fab with 57 nM targeted FAP in the presence of different concentrations of anti-EGFR GlycoMab (E: Overall A549 tumor cell kill rate due to T = 10: 1, 4 hours (see Example 8). Figure 9. Individual anti-EGFR GlycoMab (A) or Erbitux (B) (5 ng/ml or 500 ng/ml) or different concentrations of IL-2 (prolactin), 28H1 Fab-IL2-Fab or 28H1 Fab- After incubation of the combination of IL2 qm-Fab, IFN-γ released by PBMC during ADCC used A549 cells as target cells (E: T = 5: 1, 21 hours; see Example 8). Figure 10. PBMC released during non-antibody-dependent killing of A549 tumor cells after incubation with different concentrations of IL-2 (prolactin), 28H1 Fab-IL2-Fab or 28H1 Fab-IL2 qm-Fab IFN-y (E: T = 5: 1, 21 hours; see Example 8). 161561.doc •107· 201247221 Sequence Listing <11〇>Swiss Firm Rozcyliya <120> Improved Immunotherapy<130> 27306 <140> 101103935 <141> 2012-02-07 <150> EP 11153976.3 <151> 2011-02-10 <160> 156 <170> Patentln version 3.5 <210> 1 <211> 133 <212> PRT <213> Homo sapiens <400> 1

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu GIu His 15 10 15

Leu Leu Leu Asp Leu Gin Met He Leu Asn Gly He Asn Asn Tyr Lys 20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45

Lys Ala Thr Glu Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys 50 55 60

Pro Leu Glu Glu Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu 65 70 75 80

Arg Pro Arg Asp Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu 85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110

Thr lie Val Glu Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie 115 120 125 lie Ser Thr Leu Thr 130 <210> 2 <211> 133 <212> PRT <213> Homo sapiens <400> 2

Ala Pro Ala Ser Ser Ser Thr Lys Lys Thr Gin Lea Gin Leu Glu His 15 10 15

Leu Leu Leu Asp Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys 20 25 30

Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Ala Met Pro Lys 35 40 45 161561 · Sequence Listing.doc 201247221

Lys Ala Thr Glu Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys 50 55 60

Pro Leu Glu Glu Val Leu Asn Gly Ala Gin Ser Lys Asn Phe His Leu 65 70 75 80

Arg Pro Arg Asp Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu 85 90 95

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110

Thr lie Val Glu Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie 115 120 125 lie Ser Tlir Leu Thr 130 <210> 3 <211> 107 <212> PRT <213>Artificial Sequence<220><223>2B10; VL <400> 3

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Asp 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Ala Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Gly Thr Lys Val Glu lie Lys 100 . 105 <210> 4 <211> 107 <212> PRT <213>Artificial Sequence<220><223>2B10(GS); VL &lt ;400> 4

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 161561-Sequence List.doc 201247221 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Asp 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Ala Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr.Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Gly Thr Lys Val Glu lie Lys 100 105 <210> 5 <211> 121 <212> PRT <213> Artificial sequence <220><223>2B10; VH <400>

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro lie Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 012 3 Q 11 1x 1J 11 <2<2<2<2<2 6 108

PRT artificial sequence 161561-sequence table.doc 201247221 <223>2F11; VL <400> 6

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly Val Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Gin Tyr Thr Pro 85 90 95

Pro Thr Phe Gly Gin Gly Gly Thr Lys Val Glu He Lys 100 105 Column 8 τ'χ 710^人07翏37 <220><223>2F11(VI);VL<400> 7 GJu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Ττρ Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Gin Tyr Thr Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu He Lys 100 105 <210> 8 <211> 117 <212> PRT <213>Artificial Sequence-4-161561·Sequence List.doc 201247221 <220><;223>2F11; VH <400> 8

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Scr Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Scr Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Trp Arg Trp Met Met Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Oir Val Ser Ser 115 <210> 9 <211> 117 <212> PRT <213> artificial sequence <220><223>2F11(MT); VH <400>

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Trp Arg Trp Met Met Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110 161561-Sequence List.doc 201247221

Val Thr Val Ser Ser 115 <210> 10 <211> 108 <212> PRT <213> artificial sequence <220><223>3F2; VL <400>

Glu He Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Tyr Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 11 <2U> 108 <212> PRT <213>Artificial Sequence<220><223>3F2(YS); VL &lt ;400> 11

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Gly lie Met Leu Pro 85 90 95 -6- 161561 - Sequence Listing.doc 201247221

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 12 <211> 117 <212> PRT <213>Artificial Sequence<220><223>3F2; VH <400> 12

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 13 <211> 108 <212> PRT <213> artificial sequence <220><223> 3D9 > VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80 161561 · Sequence Listing.doc 201247221

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Leu lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 14 <211> 117 <212> PRT <213>Artificial Sequence<220><223> 3D9 'VH <400> 14

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Thr Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Gly Val Ser Thr Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <2I0> 15 <211> 117 <212> PRT <213> artificial sequence <220><223>3D9(TA); VH <400>

Glu Val Gin Leu Leu Glu Ser Gly Gly G】y Leu Va] G]n Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Gly Val Ser Thr Gly Ser Thr Tyr Tyr Ala Asp Ser Val 161561 · Sequence Listing.doc 201247221 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 16 <211> 108 <212> PRT <213> artificial sequence <220><223>4G8; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Arg Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45

He lie Gly Ala Ser Thr Arg Ala Thr Giy lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 ^ >>> Q1643 11 1 <2<2<2<2 17 117

PRT artificial sequence <220><223>4G8; VH <400> 17

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 161561-Sequence List.doc 201247221

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 18 <211> 108 <212> PRT <213> artificial sequence <220><223>4B3; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Asn 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Tyr lie Arg Ala Thr Gly He Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Tlir Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 19 <211> 117 <212> PRT <213>Artificial Sequence <220><223>4B3; VH <400> 19

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15 •10· 161561·Sequence List.doc 201247221

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu GIu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 SO

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 20 <211> 108 <212> PRT <213> artificial sequence <220><223>4D6; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Asn 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Gin Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 21 <211> 117 <212> PRT <213>Artificial Sequence <220>

<223>4D6; VH • 11 - 161561· Sequence Listing.doc 201247221 <400> 21

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 22 <211> 108 <212> PRT <213> artificial sequence <220><223>2C6; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Gin lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105

<210> 23 <211> 117 <212> PRT -12- 161561-sequence table.doc 201247221 <213> artificial sequence <220><223>2C6; VH <400>

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Ala Gly Tyr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110 val U Ser Ser <210> 24 <211> 108 <212> PRT <213>Artificial Sequence<220><223>5H5; VL <400> 24

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Asn Gin lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 -13- 161561 - Sequence Listing.doc 201247221 <210> 25 <211> 116 <212> PRT <213>Artificial Sequence<220>;223>5H5; VH <400> 25

Glu Val Gin Leu Leu Glu Ser Gly Gly Giy Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Thr Met Ser Trp Val Arg Arg Ser Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Gly Gly Arg Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 GJy Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80

Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95

Lys Gly Trp Phe Thr Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser 115 <210> 26 <211> 108 <212> PRT <213> artificial sequence <22Q><223>2C4; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Asn 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser lie Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Asn Gin lie Pro • 14· 161561 - Sequence Listing.doc 201247221 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 27 <211> 117 <212> PRT <213>Artificial Sequence <220>

<223>2C4; VH <400> 27

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Thr Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 28 <211> 108 <212> PRT <213> artificial sequence <220>

<223>2D9; VL <400> 28

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60 -15- 161561 - Sequence Listing.doc 201247221

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Asn Gin lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 29 <211> 117 <212> PRT <213>Artificial Sequence<220><223>2D9; VH <400> 29

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Thr Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 ^ >>> Λν 1 2 3 11 1* 11 11 <2<2<2<2 30 108

PRT artificial sequence <220><223>4B8; VL <400> 30

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 •16· 161561· Sequence Listing.doc 201247221 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr E^eu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Giu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 31 <211> 117 <212> PRT <213>Artificial Sequence<220><223>4B8; VH <400> 31

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 32 <211> 108 <212> PRT <213> artificial sequence <220><223>7A1; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30 -17- 161561 · Sequence Listing.doc 201247221

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Gin lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 33 <211> 117 <212> PRT <213>Artificial Sequence<220><223>7A1; VH <400> 33

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 34 <211> 108 <212> PRT <213> artificial sequence <220><223>13C2; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly -18* 16156 卜 Sequence Listing.doc 201247221

15 10 IS

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Leu lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 35 <211> 117 <212> PRT <213>Artificial Sequence<220><223>13C2; VH <400> 35

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 ^ >>>& 12 3 11 11 li <2<2<2<2 36 108

PRT artificial sequence <220> 161561·sequence table.doc -19. s 201247221 <223>13E8; VL <400> 36

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Leu Asn lie Pro 85 90 95

Ser Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 37 <211> 117 <212> PRT <213>Artificial Sequence<220><223>13E8; VH <400> 37

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 38 -20· 161561 - Sequence Listing.doc 201247221 <211> 108 <212> PRT <213>Artificial Sequence<220><223>14C10; VL <400> 38

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly His lie He Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 39 <211> 117 <212> PRT <213>Artificial Sequence <220><223>14C10; VH <400> 39

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Ala Trp Met Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110 -21 · 161561 - Sequence Listing.doc 201247221

Val Thr Val Ser Ser 115 <210> 40 <211> 108 <212> PRT <213> artificial sequence <22Q><223>17A11; VL <400> 40

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly He Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Leu Asn lie Pro 85 90 95

Ser Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 41 <211> 117 <212> PRT <213> Artificial Sequence <220>

<223>17A11; VH <400> 41

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 • 22- 161561 · Sequence Listing.doc 201247221

Ala Lys Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 42 <211> 108 <212> PRT <213> artificial sequence <220><223>19G1; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly He Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 43 <211> 117 <212> PRT <213>Artificial Sequence<220><223>19G1; VH <400> 43

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Ser Ser Gly Gly Leu Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 -23- 161561 - Sequence Listing.doc 201247221

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 44 <211> 108 <212> PRT <213> artificial sequence <220><223>20G8; VL <400> 44

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 45 <211> 117 <212> PRT <213>Artificial Sequence<220><223>20G8; VH <400> 45

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala He He Gly Ser Gly Ser Arg Thr Tyr Tyr Ala Asp Ser Val -24- 161561-Sequence List.doc 201247221 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Lea Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Giy Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 46 <211> 108 <212> PRT <213> artificial sequence <220><223>4B9; VL <400> 46

Glu He Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu He Lys 100 105 <210> 47 <211>117 <212> PRT <213>Artificial Sequence<220><223>4B9; VH <400> 47

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 -25- 161561 - Sequence Listing.doc 201247221

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Gly Ser Gly Ala Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 48 <211> 108 <212> PRT <213> artificial sequence <220><223>5B8; VL <400> 48

Glu He Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 49 <211> 117 <212> PRT <213>Artificial Sequence<220><223>5B8; VH <400> 49

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15 -26- 161561 - Sequence Listing.doc 201247221

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Trp Gly Gly Gly Arg Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

50th <211>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 51 <211> 117 <212> PRT <213> Artificial Sequence <220>

<223>5F1; VH • 27· 161561 - Sequence Listing. doc 3 201247221 <400> 51

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Ser Ser Gly Ala Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 52 <211> 108 <212> PRT <213> artificial sequence <220><223>14B3; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105

<210> 53 <211> 117 <212> PRT -28- 161561-sequence table, doc 201247221 <213> artificial sequence <220>

<223>14B3; VH <400> 53

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala He Leu Ala Ser Gly Ala lie Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 54 <211> 108 <212> PRT <213> artificial sequence <220><223>16F1; VL <400> 54

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly He Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 -29- 161561 - Sequence Listing.doc 201247221 <210> 55 <211> 117 <212> PRT <213>Artificial Sequence<220>;223>16F1; VH <400> 55

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Gly lie lie Gly Ser Gly Gly lie Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn TTir Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 56 <211> 108 <212> PRT <213> artificial sequence <220><223>16F8; VL <400> 56

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45

He Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Va] Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro • 30· 161561 - Sequence Listing.doc 201247221 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu He Lys 100 105 <210> 57 <211> 117 <212> PRT <213>Artificial Sequence<220><223>16F8; VH <400> 57

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Leu Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 58 <211> 108 <212> PRT <213> artificial sequence <220><223>03C9; VL <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60 • 31 · 161561 - Sequence Listing.doc 201247221

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 59 <211> 117 <212> PRT <213>Artificial Sequence<220><223>03C9; VH <400> 59

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30

Ala Met Ser Trp Val Arg Gin Ser Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Gly Ser Gly Ser Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

60th <211>

Glu lie Val Leu Hr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 -32- 161561 - Sequence Listing.doc 201247221 lie Asn Val Gly Ser Arg Arg Ala Thr Gly Thr Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Ala lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 61 <211> 117 <212> PRT <213>Artificial Sequence<220><223>02D7; VH <400> 61

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

62 lt < 211 &gt

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Arg Ser 20 25 30 •33- 161561-sequence table.doc 201247221

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie lie Gly Ala Ser Thr Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 fe> Λ > Q1^3 11 11 1A <2<2<2<2 63 116 PRT Artificial Sequence <220><223>28H1; VH <400> 63 Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Trp Ala Ser Gly Glu Gin Tyr Tyr Ala Asp Ser Val Lys 50 55 60

Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80

Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95

Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser 115 ^ >>>&123 11 11 11 11 64 108 PRT Artificial Sequence <220><223>22A3; VL <400> 64 Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 161561-Sequence List_doc .34· 201247221 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Tiir He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 65 <211> 117 <212> PRT <213>Artificial Sequence<220><223>22A3; VH <400> 65

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Gly Ser Gly Ser lie Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 66 <211> 108 <212> PRT <213> artificial sequence <220> -35 - 161561 - Sequence Listing.doc 201247221 <223>29B11; VL <400> 66

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Thr Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 67 <211> 117 <212> PRT <213>Artificial Sequence<220><223>29B11; VH <400> 67

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Gly Ser Gly Gly lie Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser 115 <210> 68 -36 - 161561 - Sequence Listing.doc 201247221 <211> 108 <212> PRT <213> Artificial Sequence <220>

<223>23C10; VL <400> 68

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Lea Ser Leu Ser Pro Gly 1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Arg Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie lie Gly Ala Ser Thr Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 69 <211> 117 <212> PRT <213>Artificial Sequence <220>

<223>23C10; VH <400> 69

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Ser 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Thr Asn Gly Asn Tyr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110 -37- 161561-sequence table.doc 201247221

Val Thr Val Ser Ser 115 <210> 70 <211> 107 <212> PRT <213> artificial sequence <220><223>2B10_C3B6; VL <400> 70

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Asp 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Ala Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly TTir Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 71 <211> 121 <212> PRT <213> Artificial Sequence <220>

<223>2B10_C3B6; VH <400> 71

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Ala lie lie Pro lie Leu Gly lie Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr He Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 -38- 161561 · Sequence Listing.doc 201247221

Ala Arg Leu Tyr Giy Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr TTir Val Thr Val Ser Ser 115 120 <210> 72 <211> 107 <212> PRT <213> artificial sequence <220><223>2B10.6A12; VL <400>

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Asp 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Ala Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 73 <211> 121 <212> PRT <213> artificial sequence <220><223>2B10_6A12; VH <400> 73

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Val lie lie Pro lie Leu Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 •39· 161561·Sequence List.doc 201247221

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 74 <211> 107 <212> PRT <213> artificial sequence <220><223>2B10.C3A6; VL <400>

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Val 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Asp Ser Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys 100 105 <210> 75 <211> 121 <212> PRT <213> Artificial sequence <220> <223>2B10_GA6; VH <400>

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro He Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe _40· 16156丨-Sequence List.doc 201247221 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 76 <211> 107 <212> PRT <213> artificial sequence <220><223>2B10eDlA2.wt; VL <400> 76

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Val 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Asp Ala Tyr Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu He Lys 100 105 <210> 77 <211> 121 <212> PRT <213>•Artificial sequence<220><223>2B10_DlA2_wt; VH <400> 77

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 -41 - 161561 - Sequence Listing.doc 201247221

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro lie Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 78 <211> 107 <212> PRT <213> artificial sequence <220><223>2B10_D1A2_VD; VL <400>

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Asp 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45·

Tyr Asp Ala Tyr Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Gly Thr Lys Val Glu lie Lys 100 105 <210> 79 <211> 121 <212> PRT <213>Artificial sequence <220><223>2B10_D1A2_VD; VH <400>

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15 • 42· 161561 · Sequence Listing.doc 201247221

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro lie Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 80 <211> 107 <212> PRT <213> artificial sequence <220><223>2B10.07D8; VL <400>

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Ser lie Arg Asn Val 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Asp Val Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu He Lys 100 105 <210> 81 <211> 121 <212> PRT <213> Artificial Sequence <220>

<223>2B10_O7D8; VH-43-161561·Sequence list.doc 201247221 <400> 81

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro lie Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe . 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110 G]n G丨y Thr Thr Va丨Thr Val Ser Ser 115 120 <210> 82 <211> 107 <212> PRT <213>Artificial sequence <220><223>2B10.01F7; VL <400> 82

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Val 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Asp Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu He Lys 100 105

<210> 83 <211> 121 <212> PRT 44·161561-sequence table.doc 201247221 <213> artificial sequence <220><223>2B10.01F7; VH <400> 83

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro lie Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 84 <211> 107 <212> PRT <213> artificial sequence <220><223>2B10.6H10; VL <400>

Asp He Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Val 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Gin Ala Ala Thr Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Gly Thr Lys Val Glu lie Lys 100 105 -45· 161561-Sequence List_doc 201247221 <210> 85 <211> 121 <212> PRT <213>Artificial Sequence<220><223>2B10_6H10; VH <400> 85

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro lie Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 86 <211> 122 <212> PRT <213> Artificial Sequence <220><223>MHLG1; VH <400>

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30

Trp Met Asn Trp Val Arg Gin Ala Pro Giy Lys Gly Leu Glu Trp Val 35 40 45

Ala Glu lie Arg Leu Lys Ser Asn Asn Phe Gly Arg Tyr Tyr Ala Ala 50 55 60

Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80

Leu Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr • 46· 161561 - Sequence Listing.doc 201247221 85 90 95

Tyr Cys Thr Thr Tyr Gly Asn Tyr Val Gly His Tyr Phe Asp His Trp 100 105 110

GL < 211 > 109 &lt

Asp He Gin Leu Thr Gin Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Asn Val Asp Thr Asn 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Pro Leu lie 35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Tyr Asn Ser Tyr Pro Leu 85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg Thr 100 105 <210> 88 <211> 122 <212> PRT <213>Artificial Sequence<220><223>MHLG; VH <400&gt ; 88

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30

Trp Met Asn Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ala Glu lie Arg Leu Lys Ser Asn Asn Phe Gly Arg Tyr Tyr Ala Ala 50 55 60 -47· 161561 - Sequence Listing _doc 201247221

Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80

Leu Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95

Tyr Cys Thr Thr Tyr Gly Asn Tyr Val Gly His Tyr Phe Asp His Trp 100 105 110

GL < 211 > 109 &lt

Asp lie Gin Leu Thr Gin Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Asn Val Asp Thr Asn 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu lie 35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe. Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Tyr Asn Ser Tyr Pro Leu 85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg Thr 100 105 <210> 90 <211> 109 <212> PRT <213>Artificial Sequence<220><223>K7; VL <400&gt ; 90

Asp lie Gin Leu Thr Gin Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Asn Val Asp Thr Asn 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Pro Leu lie 35 40 45 -48- i6156丨 · Sequence Listing.doc 201247221

Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr He Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Tyr Asn Ser Tyr Pro Leu 85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg Thr 100 105 <210> 91 <211> 603 <212> PRT <213>Artificial Sequence <220><223> Fab heavy chain - C125A variant of IL2 - Fab heavy chain derived from L19 monoclonal antibody <400>

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30

Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 1.05 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 •49· 161561·Sequence List.doc 201247221

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205

Lys Val Asp Lys Lys Va】Glu Pro Lys Ser Cys Asp Ser Gly Gly Gly 210 215 220

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser Ser 225 230 235 240

Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp Leu 245 250 255

Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu Thr 260 265 270

Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu 275 280 285

Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val 290 295 300

Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu 305 310 315 320 lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Giy Ser Glu Thr 325 330 335

Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu Phe 340 345 350

Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu Thr 355 360 365

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu 370 375 380

Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser 385 390 395 400

Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe Ser 405 410 415

Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 420 425 430

Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val Lys 435 440 445

Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 450 455 460

Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 465 470 475 480

Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr -50· 161561 · Sequence Listing.doc 201247221 485 490 495

Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 500 505 510

Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 515 520 525

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 530 535 540

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly 545 550 555 560

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 565 570 575

Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 580 585 590

Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 <210> 92 <211> 215 <212> PRT <213>Artificial Sequence <220><223> Fab derived from L19 monoclonal antibody Light chain <400> 92

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Tyr Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Thr Gly Arg lie Pro 85 90 95

Pro Tlir Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala 100 105 110

Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser 115 120 125

Glv Thr Ala Ser Val Vals Leu Leu Asn Asn Phe Tyr Pro Arg Glu 7 130 135 140 -51 - 161561 - Sequence Listing doc 201247221

Ala Lys Val Gin Trp Lys Val Asp Asn AJa Leu Gin Ser Gly Asn Ser 145 150 155 160

Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190

Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys 195 200 205

Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 93 <211> 382 <212> PRT <213>Artificial sequence <220><223> scFv-octaamino group derived from L19 monoclonal antibody Acid linker - C125A variant of IL2 <400> 93

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30

Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie SeT Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser Ser Gly Gly Ser Gly Gly Ala Ser Glu lie Val Leu 115 120 125

Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr 130 135 140

Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Serr Leu Ala Trp 145 150 155 160

Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie Tyr Tyr Ala 165 170 175 • 52· 161561 · Sequence Listing _doc 201247221

Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser Gly Ser Gly Ser 180 185 190

Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu Pro Glu Asp Phe 195 200 . 205

Ala Val Tyr Tyr Cys Gin Gin Thr Gly Arg lie Pro Pro Thr Phe Gly 210 . 215 220

Gin Gly Thr Lys Val Glu He Ser Val Leu Ser Ser Ser Ser Gly Ser 225 230 235 240

Ser Ser Ser Gly Ser Ser Ser Ser Gly Ala Pro Thr Ser Ser Ser Thr 245 250 255

Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp Leu Gin Met 260 265 270 lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met 275 280 285

Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His 290 295 300

Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn 305 310 315 320

Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu lie Ser 325 330 335

Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe 340 345 350

Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu Phe Leu Asn 355 360 365

Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu Thr 370 375 380 <210> 94 <211> 377 <212> PRT <213>Artificial sequence <220><223> F16-dual function Antibody-IL2 Protein <400> 94

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30

Gly Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val -53· 161561 - Sequence Listing.doc 201247221 35 40 45

Ser Ala He Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Ala His Asn Ala Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser Ala Ser Gly Gly Ser Ser Glu Leu Thr Gin Asp Pro 115 120 125

Ala Val Ser Val Ala Leu Gly Gin Thr Val Arg lie Thr Cys Gin Gly 130 135 140

Asp Ser Leu Arg Ser Tyr Tyr Ala Ser Trp Tyr Gin Gin Lys Pro Gly 145 150 155 160

Gin Ala Pro Val Leu Val lie Tyr Gly Lys Asn Asn Arg Pro Ser Gly 165 170 175 lie Pro Asp Arg Phe Ser Gly Ser Ser Ser Gly Asn Thr Ala Ser Leu 180 185 190

Thr He Thr Gly Ala Gin Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asn 195 200 205

Ser Ser Val Tyr Thr Met Pro Pro Val Val Phe Gly Gly Gly Thr Lys 210 215 220

Leu Thr Val Leu Gly Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ser 225 230 235 240

Ser Ser Ser Gly Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gin Leu 245 250 255

Gin Leu Glu His Leu Leu Leu Asp Leu Gin Met lie Leu Asn Gly lie 260 265 270

Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe 275 280 285

Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gin Cys Leu Glu 290 295 300

Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gin Ser Lys 305 310 315 320

Asn Phe His Leu Arg Pro Arg Asp Leu lie Ser Asn lie Asn Val lie 325 330 335 •54· 161561·Sequence List.doc 201247221

Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys GLu Tyr Ala 340 345 350

Asp Glu Thr Ala Thr lie Val Glu Phe Leu Asn Arg Trp lie Thr Phe 355 360 365

Ala Gin Ser lie lie Ser Thr Leu Thr 370 375 <210> 95 <211> 641 <212> PRT <213>Artificial sequence <220><223> scFv-IL2-scFv (F16, protein) <400> 95

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30

Gly Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Ala His Asn Ala Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Arg Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 115 120 125

Gly Gly Ser Ser Glu Leu Thr Gin Asp Pro Ala Val Ser Val Ala Leu 130 135 140

Gly Gin Thr Val Arg lie Thr Cys Gin Gly Asp Ser Leu Arg Ser Tyr 145 150 155 160

Tyr Ala Ser Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu Val 165 170 175 lie Tyr Gly Lys Asn Asn Arg Pro Ser Gly He Pro Asp Arg Phe Ser 180 185 190

Gly Ser Ser Ser Gly Asn Thr Ala Ser Leu Thr lie Thr Gly Ala Gin 195 200 205

Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Ser Val Tyr Thr Met -55- 161561-Sequence List.doc 201247221 210 215 220

Pro Pro Val 225

Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ser 230 235 240

Ser Ser Ser

Gly Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ala Pro 245 250 255

Thr Ser Ser

Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu 260 265 270

Leu Asp Leu 275

Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro 280 285

Lys Leu Thr 290

Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala 295 300

Thr Glu Leu 305

Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu 310 315 320

Glu Glu Val

Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro 325 330 335

Arg Asp Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly 340 345 350

Ser Glu Thr 355

Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie 360 365

Val Glu Phe 370

Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser 375 380

Thr Leu Thr 385

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 390 395 400

Gly Gly Ser

Ser Glu Leu Thr Gin Asp Pro Ala Val Ser Val Ala Leu 405 410 415

Gly Gin Thr

Val Arg lie Thr Cys Gin Gly Asp Ser Leu Arg Ser Tyr 420 425 430

Tyr Ala Ser 435

Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu Val 440 445 lie Tyr Gly 450

Lys Asn Asn Arg Pro Ser Gly lie Pro Asp Arg Phe Ser 455 460

Gly Ser Ser 465

Ser Gly Asn Thr Ala Ser Leu Thr lie Thr Gly Ala Gin 470 475 480

Ala Glu Asp

Glu Ala Asp Tyr Tyr Cys Asn Ser Ser Val Tyr Thr Met 485 490 495

Pro Pro Val

Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Ser 500 505 510 161561-Sequence List.doc •56· 201247221

Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Ser Glu Val 515 520 525

Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser Leu 530 535 540

Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr Gly Met 545 550 555 560

Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ala 565 570 575 lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 580 585 590

Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gin 595 600 605

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys 610 615 620

Ala His Asn Ala Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val 625 630 635 640

Ser <210> 96 <211> 603 <212> PRT < 213 > artificial sequence <220><223> Fab-IL2-Fab (F16, heavy chain cytokine fusion construct, protein) <400> 96

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr 20 25 30

Gly Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Ala His Asn Ala Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala • 57- 161561 - Sequence Listing.doc 201247221 115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Ser Ser Ser 210 215 220

Gly Ser Ser Ser Ser Gly Ser Ser Ser Ser Gly Ala Pro Thr Ser Ser 225 230 235 240

Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp Leu 245 250 255

Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu Thr 260 265 270

Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu 275 280 285

Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val 290 295 300

Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu 305 310 315 320 lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu Thr 325 330 335

Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu Phe 340 345 350

Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu Thr 355 360 365

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu 370 375 380

Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val G]n Pro Gly Gly Ser 385 390 395 400

Leu Arg. Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr Gly 405 410 415 • 58* 161561 · Sequence Listing.doc 201247221

Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 420 425 430

Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 435 440 445

Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 450 455 460

Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 465 470 475 480

Lys Ala His Asa Ala Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr 485 490 495

Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 500 505 510 .

Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 515 520 525

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 530 535 540

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly 545 550 555 560

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 565 570 575

Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 580 585 590

Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 <210> 97 <211> 214 <212> PRT <213>Artificial Sequence <220><223> F16, Light Chain, Protein <400> 97

Ser Ser Glu Leu Thr Gin Asp Pro Ala Val Ser Val Ala Leu Gly Gin 15 10 15

Thr Val Arg lie Thr Cys Gin Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30

Ser Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu Val lie Tyr 35 40 45

Gly Lys Asn Asn Arg Pro Ser Gly lie Pro Asp Arg Phe Ser Gly Ser 50 55 60

Ser Ser Gly Asn Thr Ala Ser Leu Thr lie Thr Gly Ala Gin Ala Glu -59- 16]56]-Sequence List.doc 201247221 65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Ser Val Tyr Thr Met Pro Pro 85 90 95

Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gin Pro Lys 100 105 110

Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gin 115 120 125

Ala Asn Lys Ala Thr Leu Va】Cys Leu lie Ser Asp Phe Tyr Pro Gly 130 135 140

Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly 145 150 155 160

Val Glu Thr Thr Thr Pro Ser Lys Gin Ser Asn Asn Lys Tyr Ala Ala 165 170 175

Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gin Trp Lys Ser His Arg Ser 180 185 190

Tyr Ser Cys Gin Val Thr His Glu Gly Ser TTir Val Glu Lys Thr Val 195 200 205

Ala Pro Thr Glu Cys Ser 210 <210> 98 <211> 991 <212> PRT <213>Artificial sequence<220><223> Fab-DL12-Fab, L19 antibody, murine scIL12, protein <400> 98

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Tlir Phe Ser Ser Phe 20 25 30

Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ser He Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110 -60- 161561 - Sequence Listing.doc 201247221

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly TTir Ala Ala Leu Gly Cys Leu 130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly Gly 210 215 220

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Met Trp Glu Leu 225 230 235 240

Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr Pro Asp Ala Pro 245 250 255

Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu Glu Asp Asp lie 260 265 270

Thr Trp Thr Ser Asp Gin Arg His Gly Val lie Gly Ser Gly Lys Thr 275 280 285

Leu Thr lie Thr Val Lys Glu Phe Leu Asp Ala Gly Gin Tyr Thr Cys 290 295 300

His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu Leu Leu His Lys 305 310 315 320

Lys Glu Asn Gly lie Trp Ser Thr Glu lie Leu Lys Asn Phe Lys Asn 325 330 335

Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser Gly Arg Phe Thr 340 345 350

Cys Ser Trp Leu Val Gin Arg Asn Met Asp Leu Lys Phe Asn lie Lys 355 360 365

Ser Ser Ser Ser Pro Pro Asp Ser Arg Ala Val Thr Cys Gly Met Ala 370 375 380

Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gin Arg Asp Tyr Glu Lys 385 390 395 400

Tyr Ser Val Ser Cys Gin Glu Asp Val Thr Cys Pro Thr Ala Glu Glu • 61 · 161561 · Sequence Listing.doc 201247221 405 410 415

Thr Leu Pro lie Glu Leu Ala Leu Glu Ala Arg Gin Gin Asn Lys Tyr 420 425 430

Glu Asn Tyr Set Thr Ser Phe Phe lie Arg Asp lie lie Lys Pro Asp 435 440 445

Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn Ser Gin Val Glu 450 455 460

Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro Arg Ser Tyr Phe 465 470 475 480

Ser Leu Lys Phe Phe Val Arg lie Gin Arg Lys Lys Glu Lys Met Lys 485 490 495

Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe Phe Val Glu Lys 500 505 510

Thr Ser Thr Glu Val Gin Cys Lys Gly Gly Asn Val Cys Val Gin Ala 515 520 525

Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp Ala Cys Val Pro 530 535 540

Cys Arg Val Arg Ser Gly Gly Asp Gly Ser Gly Gly Gly Gly Ser Gly 545 550 555 560

Gly Gly Gly Ser Arg Val lie Pro Val Ser Gly Pro Ala Arg Cys Leu 565 570 575

Ser Gin Ser Arg Asn Leu Lau Lys Thr Thr Asp Asp Met Val Lys Thr 580 585 590

Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp lie Asp 595 600 605

His Glu Asp lie Thr Arg Asp Gin Thr Ser Thr Leu Lys Thr Cys Leu 610 615 620

Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr 625 630 635 640

Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin Lys Thr Ser Leu 645 650 655

Met Met Thr Leu Cys Leu Gly Ser lie Tyr Glu Asp Leu Lys Met Tyr 660 665 670

Gin Thr Glu Phe Gin Ala lie Asn Ala Ala Leu Gin Asn His Asn His 675 680 685

Gin Gin lie lie Leu Asp Lys Gly Met Leu Val Ala lie Asp Glu Leu 690 695 700 -62- 16156 丨 · Sequence Listing.doc 201247221

Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gin Lys Pro Pro 705 710 715 720

Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys lie Leu 725 730 735

Leu His Ala Phe Ser Thr Arg Val Val Thr lie Asn Arg Val Met Gly 740 745 750

Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 755 760 765

Gly Gly Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin 770 775 780

Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 785 790 795 800

Ser Ser Phe Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu 805 810 815

Glu Trp Val Ser Ser lie Ser Gly Ser Ser Gly Thr Thr Tyr Tyr Ala 820 825 830

Asp Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn 835 840 845

Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 850 855 860

Tyr Tyr Cys Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly 865 870 875 880

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 885 890 895

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 900 90S 910

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 915 920 925

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 930 935 940

Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 945 950 955 960

Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro 965 970 975

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 980 985 990

<2I0> 99 <211> 987 <212> PRT • 63-161561-sequence table.doc 201247221 <213>Artificial sequence<220><223> Fab-IL12-FabL19 antibody, human SCIL12, protein <400> 99

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30

Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ser lie Arg Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly Gly 210 215 220

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly lie Trp Glu Leu Lys 225 230 235 240

Lys Asp Val Tyr Va丨 Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly 245 250 255

Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu GJu Asp Gly lie Thr 260 265 270 • 64- 161561 · Sequence Listing.doc 201247221

Trp Thr Leu Asp Gin Ser Ser GIu Val Leu Gly Ser Gly Lys Thr Leu 275 280 285

Thr lie Gin Val Lys Glu Phe Gly Asp Ala Gly Gin Tyr Thr Cys His 290 295 300

Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys Lys 305 310 315 320

Glu Asp Gly lie Trp Ser Thr Asp lie Leu Lys Asp Gin Lys Glu Pro 325 330 335

Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg 340 345 350

Phe Thr Cys Trp Trp Leu Thr Thr lie Ser Thr Asp Leu Thr Phe Ser 355 360 365

Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gin Gly Val Thr Cys Gly 370 375 380

Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr 385 390 395 400

Glu Tyr Ser Val Glu Cys Gin Glu Asp Ser Ala Cys Pro Ala Ala Glu 405 410 415

Glu Ser Leu Pro lie Glu Val Met Val Asp Ala Val His Lys Leu Lys 420 425 430

Tyr Glu Asn Tyr Thr Ser Ser Phe Phe lie Arg Asp lie lie Lys Pro 435 440 445

Asp Pro Pro Lys Asn Leu Gin Leu Lys Pro Leu Lys Asn Ser Arg Gin 450 455 460

Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser 465 470 475 480

Tyr Phe Ser Leu Thr Phe Cys Val Gin Val Gin Gly Lys Ser Lys Arg 485 490 495

Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val lie 500 505 510

Cys Arg Lys Asn Ala Ser He Ser Val Arg Ala Gin Asp Arg Tyr Tyr 515 520 525

Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly Gly 530 535 540

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Arg Asn Leu Pro 545 550 555 560

Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu His His Ser Gin 565 570 575 • 65- 161561 - Sequence Listing.doc 201247221

Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gin Lys Ala Arg Gin Thr 580 585 590

Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu lie Asp His Glu Asp lie 595 600 605

Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu Pro Leu Glu Leu 610 615 620

Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr Ser Phe lie Thr 625 630 635 640

Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe Met Met Ala Leu 645 650 655

Cys Leu Ser Ser lie Tyr Glu Asp Leu Lys Met Tyr Gin Val Glu Phe 660 665 670

Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys Arg Gin lie Phe 675 680 685

Leu Asp Gin Asn Met Leu Ala Val lie Asp Glu Leu Met Gin Ala Leu 690 695 700

Asn Phe Asn Ser Glu Thr Val Pro Gin Lys Ser Ser Leu Glu Glu Pro 705 710 715 720

Asp Phe Tyr Lys Thr Lys lie Lys Leu Cys lie Leu Leu His Ala Phe 725 730 735

Arg lie Arg Ala Val Thr lie Asp Arg Val Met Ser Tyr Leu Asn Ala 740 745 .750

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu 755 760 765

Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser 770 775 780

Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe Ser 785 790 795 800

Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 805 810 815

Ser lie Arg Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val Lys 820 825 830

Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 835 840 845

Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 850 855 860

Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr 865 870 875 880 • 66· 161561 · Sequence Listing.doc 201247221

Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 885 890 895

Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 900 905 910

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 915 920 925

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly 930 935 940

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 945 950 955 960

Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 965 970 975

Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 980 985 <210> 100 <211> 597 <212> PRT <213>Artificial Sequence<220><223> Fab-GMCSF-Fab, L19 Antibody , human GM-CSF, protein <400> 100

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30

Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ser lie Arg Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 • 67· 161561 · Sequence Listing.doc 201247221

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly Gly 210 215 220

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Ala Arg Ser 225 230 235 240

Pro Ser Pro Ser Thr Gin Pro Trp Glu His Val Asn Ala lie Gin Glu 245 250 255

Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met Asn 260 265 270

Glu Thr Val Glu Val lie Ser Glu Met Phe Asp Leu Gin Glu Pro Thr 275 280 285

Cys Leu Gin Thr Arg Leu Glu Leu Tyr Lys Gin Gly Leu Arg Gly Ser 290 295 300

Leu Thr Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr Lys 305 310 315 320

Gin His Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gin lie lie 325 330 335

Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val lie 340 345 350

Pro Phe Asp Cys Trp Glu Pro Val Gin Glu Ser Gly Gly Gly Gly Ser 355 360 365

Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gin Leu Leu Glu Ser 370 375 380

Gly Gly Gly Leu Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala 385 390 395 400

Ala Ser Gly Phe Thr Phe Ser Ser Phe Ser Met Ser Trp Val Arg Gin 405 410 415

Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser lie Arg Gly Ser Ser 420 425 430

Gly Thr Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr He Ser 435 440 445 • 68· 161561 - Sequence Listing.doc 201247221

Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg 450 455 460

Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Pro Phe Pro Tyr Phe 465 470 475 480

Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 485 490 495

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 500 505 510

Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 515 520 525

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Lea Thr Ser Gly Val His 530 535 540

Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser 545 550 555 560

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys 565 570 575

Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 580 585 590

Pro Lys Ser Cys Asp 595 <210> 101 <211> 636 <212> PRT <213>Artificial sequence <220><223> Fab-IFNa2-Fab, L19 antibody, protein <400>

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 20 25 30

Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ser lie Arg Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 -69· 161561 - Sequence Listing.doc 201247221

Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly Gly 210 215 220

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Cys Asp Leu Pro Gin 225 230 235 240

Thr His Ser Leu Gly Asn Arg Arg Ala Leu He Leu Leu Ala Gin Met 245 250 255

Arg Arg lie Ser Pro Phe Ser Cys Leu Lys Asp Arg His Asp Phe Gly 260 265 270

Phe Pro Gin Glu Glu Phe Asp Gly Asn Gin Phe Gin Lys Ala Gin Ala 275 280 285 lie Ser Val Leu His Glu Met lie Gin Gin Thr Phe Asn Leu Phe Ser 290 295 300

Thr Lys Asp Ser Ser Ala Ala Trp Asp Glu Ser Leu Leu Glu Lys Phe 305 310 315 320

Tyr Thr Glu Leu Tyr Gin Gin Leu Asn Asp Leu Glu Ala Cys Val lie 325 330 335

Gin Glu Val Gly Val Glu Glu Thr Pro Leu Met Asn Val Asp Ser lie 340 345 350

Leu Ala Val Lys Lys Tyr Phe Gin Arg lie Thr Leu Tyr Leu Thr Glu 355 360 365

Lys Lys Tyr Ser Pro Cys Ala Trp Glu Val Val Arg Ala Glu lie Met 370 375 380

Arg Ser Phe Ser Leu Ser Thr Asn Leu Gin Glu Arg Leu Arg Arg Lys 385 390 395 400 -70· 161561 · Sequence Listing.doc 201247221

Glu Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 405 410 415

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 420 425 430

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe 435 440 445

Ser Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 450 455 460

Ser Ser lie Arg Gly Ser Ser Gly Thr Thr Tyr Tyr Ala Asp Ser Val 465 470 475 480

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 485 490 495

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 500 505 510

Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 515 520 525

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 530 535 540

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 545 550 555 560

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 565 570 575

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 580 585 590

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 595 600 605

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 610 615 620

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 625 630 635 <210> 102 <211> 605 <212> PRT <213>Artificial Sequence<220><223> 3F2Fab-IL2-Fab ( Heavy chain cytokine fusion construct) <400> 102

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15 -71 - 161561 · Sequence Listing.doc 201247221

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp 305 310 315 320 • 72· 161561 - Sequence Listing.doc 201247221

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu 340 345 350

Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365

Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 370 375 380

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 420 425 430

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 465 470 475 480

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 103 73-161561-Sequence List_doc 201247221 <211> 605 <212> PRT <213>Artificial Sequence<220><223> 4G8Fab-IL2-Fab (heavy chain cytokine fusion construct) <400> 103

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Va] 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Hir Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly G】y 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270 -74- 161561-sequence table.doc 201247221

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Hie His Leu Arg Pro Arg Asp 305 310 315 320

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu 340 345 350

Phe Leu Asn Arg Trp He Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365 ^SerGlyGlyGlyGlySerGlyG, yGlyGlySerGlyGlyGlyGly

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 420 425 430

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 465 470 475 480

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser • 75- 161561 - Sequence Listing.doc 201247221 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 104 <211> 605 <212> PRT <213>Artificial Sequence<220><223> 3D9Fab-IL2-Fab (heavy chain cytokine fusion construct) <400> 104

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Tlir Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Thr Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Gly Val Ser Thr Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr He Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220 -76· 161561 · Sequence Listing.doc 201247221

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly He Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp 305 310 315 320

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu 340 345 350

Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365

Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 370 375 380

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Thr Pro Gly Lys Gly Leu Glu Trp Val 420 425 430

Ser Ala lie Gly Val Ser Thr Gly Ser Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 465 470 475 480

Ala Lys Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys -77- 161561 · Sequence Listing.doc 201247221 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 105 <211> 605 <212> PRT <213>Artificial Sequence<220><223> 2FllFab-IL2-Fab (heavy chain cytokine fusion construct) <400> 105

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Trp Arg Trp Met Met Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 · 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175 •78· 161561-sequence table.doc 201247221

Scr Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp 305 310 315 320

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu 340 345 350

Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365

Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 370 375 380

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 420 425 430

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 161561-Sequence List.doc -79-

S 201247221 465 470 475 480

Ala Lys Trp Arg Trp Met Met Phe Asp Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 106 <211> 605 <212> PRT <213>Artificial Sequence<220><223> 4B3Fab-IL2-Fab (heavy chain cytokine fusion construct) <400> 106

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala He Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 •80- 161561-sequence table.doc 201247221

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp 305 310 315 320

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu 340 345 350

Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365

Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Scr Gly Gly Gly Gly 370 375 380

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val • 81. 161561 - Sequence Listing doc 201247221 420 425 430

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 465 470 475 480

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 107 <211> 993 <212> PRT <213>Artificial Sequence<220><223> 4G8Fab-IL12-Fab (murine IL-12; heavy chain cytokine fusion construct) <400> 107

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr -82- 161561 - Sequence Listing.doc 201247221 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Scr 145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Met Trp Glu 225 230 235 240

Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr Pro Asp Ala 245 250 255

Pro Gly Glu Thr Val Asn Leu Thr Cys Asp TTir Pro Glu Glu Asp Asp 260 265 270 lie Thr Trp Thr Ser Asp Gin Arg His Gly Val lie Gly Ser Gly Lys 275 280 285

Thr Leu Thr He Thr Val Lys Glu Phe Leu Asp Ala Gly Gin Tyr Thr 290 295 300

Cys His Lys Gly Gly Glu Thr Leu Ser His Scr His Leu Leu Leu His 305 310 315 320

Lvs Lvs Glu Asn Gly lie Trp Ser Thr Glu He Leu Lys Asn Phe Lys 325 330 335

Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser Gly Arg Phe 340 345 350

Thr Cys Ser Trp Leu Val Gin Arg Asn Met Asp Leu Lys Phe Asn lie 355 360 365 •83· 161561-sequence table doc 201247221

Lys Ser Ser Ser Ser Pro Pro Asp Ser Arg Ala Val Thr Cys Gly Met 370 375 380

Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gin Arg Asp Tyr Glu 385 390 395 400

Lys Tyr Ser Val Ser Cys GJn Glu Asp Val Thr Cys Pro TTir Ala Glu 405 410 415

Glu Thr Leu Pro He Glu Leu Ala Leu Glu Ala Arg Gin Gin Asn Lys 420 425 430

Tyr Glu Asn Tyr Ser Thr Ser Phe Phe lie Arg Asp lie lie Lys Pro 435 440 445

Asp Pro Pro Lys Asn Leu Gin Met Lys Pro Leu Lys Asn Ser Gin Val 450 455 460

Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro Arg Ser Tyr 465 470 475 480

Phe Ser Leu Lys Phe Phe Val Arg lie Gin Arg Lys Lys Glu Lys Met 485 490 495

Lys Glu Thr Glu Glu Gly Cys Asn Gin Lys Gly Ala Phe Phe Val Glu 500 505 510

Lys TTir Ser Thr Glu Val Gin· Cys Lys Gly Gly Asn Val Cys Val Gin 515 520 525

Ala Gin Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp Ala Cys Val 530 535 540

Pro Cys Arg Val Arg Ser Gly Gly Asp Gly Ser Gly Gly Gly Gly Ser 545 550 555 560

Gly Gly Gly Gly Ser Arg Val lie Pro Val Ser Gly Pro Ala Arg Cys 565 570 575

Leu Ser Gin Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys 580 585 590

Thr Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp lie 595 600 605

Asp His Glu Asp lie Thr Arg Asp Gin Thr Ser Thr Leu Lys Thr Cys 610 615 620

Leu Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu 625 630 635 640

Thr Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gin Lys Thr Ser 645 650 655

Leu Met Met Thr Leu Cys Leu Gly Ser lie Tyr Glu Asp Leu Lys Met 660 665 670 •84- 161561 · Sequence Listing.doc 201247221

Tyr Gin Thr Glu Phe Gin Ala lie Asn Ala Ala Leu Gin Asn His Asn 675 680 685

His Gin Gin lie lie Leu Asp Lys Gly Met Leu Val Ala lie Asp Glu 690 695 700

Leu Met Gin Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gin Lys Pro 705 710 715 720

Pro Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys lie 725 730 735

Leu Leu His Ala Phe Ser Thr Arg Val Val Thr lie Asn Arg Val Met 740 745 750

Gly Tyr Leu Ser Ser Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 755 760 765

Gly Gly Gly Gly Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val 770 775 780

Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 785 790 795 800

Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly 805 810 815

Leu Glu Trp Val Ser Ala lie Ser Gly Ser Gly Gly Ser Thr Tyr Tyr 820 825 830

Ala Asp Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys 835 840 845

Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 850 855 860

Val Tyr Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly 865 870 875 880

Gin Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 885 890 895

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 900 905 910

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 915 920 925

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 930 935 940

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Tlir Val 945 950 955 960

Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 965 970 975 -85 - 161561 · Sequence Listing.doc 201247221

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 980 985 990

Asp <210> 108 <211> 603 <212> PRT < 213 > artificial sequence <220><223> 28HlFab-IL2-Fab (heavy chain cytokine fusion construct) <400>

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie Trp Ala Ser Gly Glu Gin Tyr Tyr Ala Asp Ser Val Lys 50 55 60

Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80

Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95

Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val 100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125

Pro Ser Ser Lys Ser TTir Ser Gly Gly Ήιγ Ala Ala Leu Gly Cys Leu 130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser 165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190

Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly Gly 210 215 220 -86- 161561 - Sequence Listing.doc 201247221

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser Ser 225 230 235 240

Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp Leu .245 250 255

Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu Thr 260 265 270

Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu Leu 275 280 285

Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu Val 290 295 300

Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp Leu 305 310 315 320 lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu Thr 325 330 335

Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu Phe 340 345 350

Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu Thr 355 360 365

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu 370 375 380

Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser 385 390 395 400

Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His Ala 405 410 415

Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Ser 420 425 430

Ala lie Trp Ala Ser Gly Glu Gin Tyr Tyr Ala Asp Ser Val Lys Gly 435 440 445

Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gin 450 455 460

Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys 465 470 475 480

Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr 485 490 495

Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 500 505 510

Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 515 520 525 -87- 161561-sequence table.doc 201247221

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 530 535 540

Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly 545 550 555 560

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 565 570 575

Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 580 585 590

Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 <210> 109 <211> 605 <212> PRT <213>Artificial Sequence<223> 29B11 Fab-IL2-Fab (Heavy Chain Cytokine Fusion Construction) <400> 109

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Gly Ser Gly Gly lie Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160

Gly A]a Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175 -88 - 16156] - Sequence Listing.doc 201247221

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp 305 310 315 320

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu 340 345 350

Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365

Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 370 375 380

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 420 425 430

Ser Ala lie lie Gly Ser Gly Gly lie Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 465 470. 475 480 -89- 161561-sequence table.doc 201247221

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590 TTir Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 110 <211> 605 <212> PRT <213> Artificial sequence <220><223> 19GlFab-IL2-Fab (heavy chain cytokine fusion construct) <400>

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Ser Ser Gly Gly Leu Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 •90- 161561-sequence table.doc 201247221

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp 305 310 315 320

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr He Val Glu 340 345 350

Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365

Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly .370 375 380

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 420 425 430 •91- 161561-sequence table.doc 201247221

Ser Ala lie lie Ser Ser Gly Gly Leu Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 465 470 475 480

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 111 <211> 605 <212> PRT <213>Artificial Sequence<220><223> 20G8Fab-IL2-Fab (heavy chain cytokine fusion construct) <400> 111

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Ala lie lie Gly Ser Gly Ser Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 -92- 161561 - Sequence Listing.doc 201247221

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 100 105 110

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ser Gly Gly 210 215 220

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ala Pro Thr Ser 225 230 235 240

Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His Leu Leu Leu Asp 245 250 255

Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys Asn Pro Lys Leu 260 265 270

Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys Lys Ala Thr Glu 275 280 285

Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys Pro Leu Glu Glu 290 295 300

Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu Arg Pro Arg Asp 305 310 315 320

Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu Lys Gly Ser Glu 325 330 335

Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala Thr lie Val Glu 340 345 350

Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie lie Ser Thr Leu 355 360 365

Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 370 375 380 • 93· 161561 · Sequence Listing.doc 201247221

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 385 390 395 400

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 405 410 415

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 420 425 430

Ser Ala lie lie Gly Ser Gly Ser Arg Thr Tyr Tyr Ala Asp Ser Val 435 440 445

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 450 455 460

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 465 470 475 480

Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gin Gly Thr Leu 485 490 495

Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 500 505 510

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 515 520 525

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 530 535 540

Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser 545 550 555 560

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 565 570 575

Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn 580 585 590

Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 595 600 605 <210> 112 <211> 215 <212> PRT <213>Artificial Sequence <220><223> 3F2 Light Chain <400> 112

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val ITir Ser Ser 20 25 30 -94- 161561 · Sequence Listing.doc 201247221

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Asn Val Gly Ser Arg Arg Ala Thr Gly He Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly lie Met Leu Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala 100 105 110

Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser 115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140

Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser 145 150 155 160

Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190

Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys 195 200 205

Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 113 <211> 215 <212> PRT <213>Artificial sequence <220><223> 4G8 light chain <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Arg Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie lie Gly Ala Ser Thr Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60 -95- 161561 · Sequence Listing.doc 201247221

Gly Ser Gly Ser Gly Thr Asp Phe TTir Leu Thr He Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala 100 105 110

Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser 115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140

Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser 145 150 155 160

Gla Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190

Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys 195 200 205

Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 114 <211> 215 <212> PRT <213>Artificial Sequence <220><223> 3D9 Light Chain <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45

He Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Leu lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala • 96· 161561 - Sequence Listing.doc 201247221 100 105 110

Ala Pro Scr Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser 115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 . 135 140

Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Scr 145 150 155 160

Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175

Ser Scr Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190

Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys 195 200 205

Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 115 <211> 215 <212> PRT <213>Artificial Sequence <220><223> 2F11 Light Chain <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Scr Leu Scr Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Ser 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Ser Ser Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gly Gin Tyr Thr Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala 100 105 110

Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Scr 115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 -97- 161561-sequence table.doc 201247221

Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser 145 150 155 160

Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190

Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys 195 200 205

Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 116 <211> 215 <212> PRT <213> Artificial sequence <220><223> 4B3 light chain <400>

Glu lie Val Leu Thr Gin Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Asn 20 25 30

Tyr Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu 35 40 45 lie Tyr Gly Ala Tyr lie Arg Ala Thr Gly lie Pro Asp Arg Phe Ser 50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Arg Leu Glu 65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Gin Gin Val lie Pro 85 90 95

Pro Thr Phe Gly Gin Gly Thr Lys Va] Glu lie Lys Arg Thr Val Ala 100 105 110

Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser 115 120 125 .

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140

Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser 145 150 155 160

Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 -98- 161561 - Sequence Listing.doc 201247221

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190

Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys 195 200 205

Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 117 <211> 613 <212> PRT <213>Artificial Sequence<220><223> 2B10Fab-IL2-Fab (Heavy Chain Cytokine Fusion Construction)体) <400> 117

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala He Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Gly lie lie Pro lie Phe Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 195 200 205 •99- 161561-Sequence List.doc 201247221

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220

Asp Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 225 230 235 240

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His 245 250 255

Leu Leu Leu Asp Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr Lys 260 265 270

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 275 280 285

Lys Ala Thr Glu Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys 290 295 300

Pro Leu Glu Glu Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu 305 310 315 320

Arg Pro Arg Asp Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu 325 330 335

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 340 345 350

Thr lie Val Glu Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser lie 355 360 365 lie Ser Thr Leu Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 370 375 380

Gly Gly Gly Gly Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys 385 390 395 400

Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr 405 410 415

Phe Ser Ser Tyr Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly 420 425 430

Leu Glu Trp Met Gly Gly He lie Pto lie Phe Gly Thr Ala Asn Tyr 435 440 445

Ala Gin Lys Phe Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr 450 455 460

Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala 465 470 475 480

Val Tyr Tyr Cys Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe 485 490 495

Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 500 505 510 -100 - 161561 - Sequence Listing.doc 201247221

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 515 520 525

Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 530 535 540

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 545 550 555 560

Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser 565 570 575

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys 580 585 590

Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 595 600 605

Pro Lys Ser Cys Asp 610 <210> 118 <211> 613 <212> PRT <213>Artificial Sequence<220><223> C3B6Fab-EL2-F^) (heavy chain cytokine fusion construct) ) <400> 118

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Ala lie lie Pro lie Leu Gly lie Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125

Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 -101 - 161561 - Sequence Listing.doc 201247221

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175

Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190

Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn His 195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220

Asp Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 225 230 235 240

Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu His 245 250 255

Leu Leu Leu Asp Leu Gin Met He Leu Asn Gly lie Asn Asn Tyr Lys 260 265 270

Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 275 . 280 285

Lys Ala Thr Glu Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu Lys 290 295 300

Pro Leu Glu Glu Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His Leu 305 310 315 320

Arg Pro Arg Asp Leu lie Ser Asn lie Asn Val lie Val Leu Glu Leu 325 330 335

Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 340 345 350

Thr lie Val Glu Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser He 355 360 365 lie Ser Thr Leu Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 370 375 380

Gly Gly Gly Gly Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys 385 390 395 400

Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr 405 410 415

Phe Ser Ser Tyr Ala He Ser Trp Val Arg Gin Ala Pro Gly Gin Gly 420 425 430

Leu Glu Trp Met Gly Ala lie lie Pro lie Leu Gly lie Ala Asn Tyr 435 440 445 -102- 161561 · Sequence Listing.doc 201247221

Ala Gin Lys Phe Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr 450 455 460

Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala 465 470 475 480

Val Tyr Tyr Cys Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe 485 490 495

Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 500 505 510

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 515 520 525

Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 530 535 540

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu TTir Ser Gly Val His 545 550 555 560

Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser 565 570 575

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys 580 585 590

Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 595 600 605

Pro Lys Ser Cys Asp 610 <210> 119 <211> 613 <212> PRT <213>Artificial sequence <220><223> 6A12 Fab-IL2-Fab (heavy chain cytokine fusion construct) <400> 119

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Val lie lie Pro lie Leu Gly Thr Ala Asn Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 -103- 161561 - Sequence Listing.doc 201247221

Met Glu Leu Ser Ser Leu 85

Arg Ser Glu Asp Thr Ala 90

Val Tyr Tyr Cys 95

Ala Arg Leu Tyr Gly Tyr 100

Ala Tyr Tyr Gly Ala Phe 105

Asp Tyr Trp Gly 110

Gin Gly Thr Thr Val Thr 115

Val Ser Ser Ala Ser Thr 120

Lys Gly Pro Ser 125

Val Phe Pro Leu Ala Pro 130

Ser Ser Lys Ser Thr Ser 135 140

Gly Gly Thr Ala

Ala Leu Gly Cys Leu Val 145 150

Lys Asp Tyr Phe Pro Glu 155

Pro Val Thr Val 160

Ser Trp Asn Ser Gly Ala 165

Leu Thr Ser Gly Val His 170

Thr Phe Pro Ala 175

Val Leu Gin Ser Ser Gly 180

Leu Tyr Ser Leu Ser Ser 185

Val Val Thr Val 190

Pro Ser Ser Ser Leu Gly 195

Thr Gin Thr Tyr lie Cys 200

Asn Val Asn His 205

Lys Pro Ser Asn Thr Lys 210

Val Asp Lys Lys Val Glu 215 220

Pro Lys Ser Cys

Asp Ser Gly. Gly Gly Gly 225 230

Ser Gly Gly Gly Gly Ser 235

Gly Gly Gly Gly 240

Ala Pro Thr Ser Ser Ser 245

Thr Lys Lys Thr Gin Leu 250

Gin Leu Glu His .255

Leu Lea Leu Asp Leu Gin 260

Met lie Leu Asn Gly lie 265

Asn Asn Tyr Lys 270

Asn Pro Lys Leu Thr Arg 275

Met Leu Thr Phe Lys Phe 280

Tyr Met Pro Lys 285

Lys Ala Thr Glu Leu Lys 290

His Leu Gin Cys Leu Glu 295 300

Giu Glu Leu Lys

Pro Leu Glu Glu Val Leu 305 310

Asn Leu Ala Gin Ser Lys 315

Asn Phe His Leu 320

Arg Pro Arg Asp Leu lie 325

Ser Asn lie Asn Val lie 330

Val Leu Glu Leu 335

Lys Gly Ser Glu Thr Thr 340

Phe Met Cys Glu Tyr Ala 345

Asp Glu Thr Ala 350

Thr lie Val Glu Phe Leu 355

Asn Arg Trp lie Thr Phe 360

Ala Gin Ser lie 365 lie Ser Thr Leu Thr Ser 370

Gly Gly Gly Gly Ser Gly 375 380

Gly Gly Gly Ser • 104· 161561· Sequence Listing.doc 201247221

Gly Gly Gly Gly Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys 385 390 395 400

Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr 405 410 415

Phe Ser Ser Tyr Ala lie Ser Trp Val Arg Gin Ala Pro Gly Gin Gly 420 425 430

Leu Glu Trp Met Gly Val lie lie Pro lie Leu Gly Thr Ala Asn Tyr 435 440 445

Ala Gin Lys Phe Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr 450 455 460

Ser Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala 465 470 475 480

Val Tyr Tyr Cys Ala Arg Leu Tyr Gly Tyr Ala Tyr Tyr Gly Ala Phe 485 490 495

Asp Tyr Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 500 505 510

Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 515 520 525

Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 530 535 540

Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 545 550 555 560

Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser 565 570 575

Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys 580 585 590

Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 595 600 605

Pro Lys Ser Cys Asp 610 <210> 120 <211> 214 <212> PRT <213>Artificial Sequence<220><223> 2B10 Light Chain <400> 120 ^sp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15 -105- 161561 - Sequence Listing.doc 201247221

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Asp 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Ala Ala Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Va] Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser G] n 145 150 155 160

Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205

Phe Asn Arg Gly Glu Cys 210 b > Λ > ^ >&123& 3 11 n il 1i oz 2 ^<2 121 214

PRT artificial sequence D1A2 light chain <400> 121

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Arg Asn Asp 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Asp Ala Tyr Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly -106- 161561-Sequence List.doc 201247221 50 55 60

Gly Gly Ser Gly Thr Glu Phe Thr Leu Thr He Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95

Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala Ala loo 105 no

Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin 145 150 155 160

Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205

"health sequence <220> &lt

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Ser lie Arg Asn Val 20 25 30

Leu Gly Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Asp Val Ser Ser Leu Gin Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Gly Gly Scr Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin Asn Gly Leu Gin Pro Ala 85 90 95 -107- 161561 · Sequence Listing.doc 201247221

Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin 145 150 155 160

Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205

Phe Asn Arg Gly Glu Cys 210 <210> 123 <211> 615 <212> PRT <213>Artificial Sequence<220><223> MHLG1 Fab-IL2-Fab (heavy chain cytokine fusion construct) ) <400> 123

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30

Trp Met Asn Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ala Glu lie Arg Leu Lys Ser Asn Asn Phe Gly Arg Tyr Tyr Ala Ala 50 55 60

Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80

Leu Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95

Tyr Cys Thr Thr Tyr Gly Asn Tyr Val Gly His Tyr Phe Asp His Trp 100 105 110

Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 •108· 】6156]-Sequence List, doc 201247221

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175

Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Ήιγ 180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn 195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220

Cys Asp Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 225 230 235 240

Gly Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu 245 250 255

His Leu Leu Leu Asp Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr 260 265 270

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 275 280 285

Lys Lys Ala Thr Glu Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu 290 295 300

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His 305 310 315 320

Leu Arg Pro Arg Asp Leu lie Ser Asn lie Asn Val lie Val Leu Glu 325 330 335

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 340 345 350

Ala Thr lie Val Glu Phe Leu Asn Arg Trp He Thr Phe Ala Gin Ser 355 360 365 lie lie Ser Thr Leu Thr Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 370 375 380

Ser Gly Gly Gly Gly Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu 385 390 395 400

Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 405 410 415

Thr Phe Ser Asn Tyr Trp Met Asn Trp Val Arg Gin Ala Pro Gly Lys 420 425 430 -109- 161561-sequence table.doc 201247221

Gly Leu Glu Trp Val Ala Glu lie Arg Leu Lys Ser Asn Asn Phe Gly 435 440 445

Arg Tyr Tyr Ala Ala Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp 450 455 460

Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu 465 470 475 480

Asp Thr Ala Val Tyr Tyr Cys Thr Thr Tyr Gly Asn Tyr Val Gly His 485 490 495

Tyr Phe Asp His Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala 500 505 510

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 515 520 525

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 530 535 540

Pro Glu Pro Val Thr Va] Ser Trp Asn Ser Gly Ala Leu Tlir Ser Gly 545 550 555 560

Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu 565 570 575

Ser Ser Val Val Val Val Val Val Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser

Val Glu Pro Lys Ser Cys Asp 610 615 <210> 124 <211> 214 <212> PRT <213>Artificial Sequence <220><223> KV9 Light Chain <400> 124

Asp He Gin Leu Thr Gin Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Asn Val Asp Thr Asn 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly G】n Ala Pro Arg Pro Leu lie 35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 -110· 161561-Sequence List.doc 201247221

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Tyr Asn Ser Tyr Pro Leu 85 - 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin 145 150 155 160

Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205

Phe Asn Arg Gly Glu Cys 210 <210> 125 <211> 615 <212> PRT <213>Artificial Sequence<220><223> MHLGFab-IL2-Fab (Heavy Chain Cytokine Fusion Construct) <400> 125

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30

Trp Met Asn Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ala Glu lie Arg Leu Lys Ser Asn Asn Phe Gly Arg Tyr Tyr Ala Ala 50 55 60

Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp Asp Ser Lys Asn Thr 65 70 75 80

Leu Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95 -Ill - 161561 · Sequence Listing.doc s 201247221

Tyr Cys Thr Thr Tyr Gly Asn Tyr Val Gly His Tyr Phe Asp His Trp 100 105 110

Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175

Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys Asn Val Asn 195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220

Cys Asp Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 225 230 235 240

Gly Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gin Leu Gin Leu Glu 245 250 255

His Leu Leu Leu Asp Leu Gin Met lie Leu Asn Gly lie Asn Asn Tyr 260 265 270

Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro 275 280 285

Lys Lys Ala Thr Glu Leu Lys His Leu Gin Cys Leu Glu Glu Glu Leu 290 295 300

Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gin Ser Lys Asn Phe His 305 310 315 320

Leu Arg Pro Arg Asp Leu lie Ser Asn lie Asn Val lie Val Leu Glu 325 330 335

Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr 340 345 350

Ala Thr lie Val Glu Phe Leu Asn Arg Trp lie Thr Phe Ala Gin Ser 355 360 365 lie lie Ser Thr Leu Tlir Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 370 375 380

Ser Gly Gly Gly Gly Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu 385 390 395 400 -112· 161561 · Sequence Listing.doc 201247221

Val Gin Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 405 410 415

Thr Phe Ser Asn Tyr Trp Met Asn Trp Val Arg Gin Ala Pro Gly Lys 420 425 430

Gly Leu Glu Trp Val Ala Glu lie Arg Leu Lys Ser Asn Asn Phe Gly 435 440 445

Arg Tyr Tyr Ala Ala Ser Val Lys Gly Arg Phe Thr lie Ser Arg Asp 450 455 460

Asp Ser Lys Asn Thr Leu Tyr Leu Gin Met Asn Ser Leu Lys Thr Glu 465 470 475 480

Asp Thr Ala Val Tyr Tyr Cys Thr Thr Tyr Gly Asn Tyr Val Gly His 485 490 495

Tyr Phe Asp His Trp Gly Gin Gly Thr Thr Val Thr Val Ser Ser Ala 500 505 510

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser 515 520 525

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe 530 535 540

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly 545 550 555 560

Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu 565 570 575

Ser Ser Val Val Val Val Val Val Ser Ser Val Val Val Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser

VAL > 211 <212&gt

Asp lie Gin Leu Thr Gin Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Asn Val Asp Thr Asn 20 25 30 •113· 161561-Sequence List.doc 201247221

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu He 35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Ήιγ Gly Va】Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Tyr Asn Ser Tyr Pro Leu 85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin 145 150 155 160

Glu Ser Val Thr G】u Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val TTu Lys Ser 195 200 205

PG < 211 &lt

Asp lie Gin Leu Thr Gin Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr He Thr Cys Lys Ala Ser Gin Asn Val Asp Thr Asn 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Pro Leu lie 35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Va】Pro Ser Arg Phe Ser G】y 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro • 114- 161561 · Sequence Listing.doc 201247221 80 65 70 75

Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Tyr Asn Ser Tyr Pro Leu 85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys Arg Thr Val Ala Ala 100 105 110

Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140

Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin 145 150 155 160

Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190

Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205

"health sequence <220&gt

Gly Tyr Ala Phe Ser Tyr <210> 129 <211> 8 <212> PRT <213>Artificial sequence <220><223> Resistance 201^〇112 <400>

"health sequence <220> &lt List .doc 201247221 <400> 130

Asn Val Phe Asp Gly Tyr Trp Leu Val Tyr 1 5 10 <210> 131 <211> 16 <212> PRT <213> Artificial sequence <220><223> Anti-CD20 LCDR1 <400>

Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly lie Thr Tyr Leu Tyr 1 5 10 15 <210> 132 <211> 7 <212> PRT <213> Artificial Sequence <220><223> CD20LCDR2 <400> 132

Gin Met Ser Asn Leu Val Ser <210> 133 <211> 9 <212> PRT <213> artificial sequence <220><223> ^lCD20LCDR3 <400>

134 <211&gt

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Va] Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ala Phe Ser Tyr Ser 20 25 30

Trp lie Asn Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Arg lie Phe Pro Gly Asp Gly Asp Thr Asp Tyr Asn Gly Lys Phe 50 55 60

Lys Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 116- 161561 · Sequence Listing.doc 201247221 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Asn Val Phe Asp Gly Tyr Trp Leu Val Tyr Trp Gly Gin Gly 100 105 110

Thr Leu Val Thr Val Ser Ser 115 ^ >>> 164 3 11 1L 22^2 VC V < 135 115

PRT artificial sequence <220><223> anti 0020\^ <400> 135

Asp lie Val Met Thr Gin Thr Pro Leu Ser Leu Pro Val Thr Pro Gly 15 10 15

Glu Pro Ala Ser lie Ser Cys Arg Ser Ser Lys Ser Leu Leu His Ser 20 25 30

Asn Gly lie Thr Tyr Leu Tyr Trp Tyr Leu Gin Lys Pro Gly Gin Ser 35 40 45

Pro Gin Leu Leu lie Tyr Gin Met Ser Asn Leu Val Ser Gly Val Pro 50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys lie 65 70 75 80

Ser Arg Val Glu Aia Glu Asp Val Gly Val Tyr Tyr Cys Ala Gin Asn 85 90 95

Leu Glu Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu lie Lys 100 105 110

Arg Thr Val 115 <210> 136 <211> 5 <212> PRT <213> artificial sequence <220><223> ^.EGFRHCDR1 <400>

Asp Tyr Lys lie His <210> 137 <211> 17 <212> PRT <213> artificial sequence 117-161561-sequence table.doc 201247221 <220><223> posted GFRHCDR2 <400> Tyr Phe Asn Pro Asn Ser Gly Tyr Ser Thr Tyr Ala Gin Lys Phe Gin 15 10 15 ^ >>>& 1 2 3 11 11 11 11 5^3⁄4^ 138 11 PRT Artificial Sequence <220><223> Anti-£«11^«:0113 <400> 138 Leu Ser Pro Gly Gly Tyr Tyr Val Met Asp Ala 1 5 10 <210> 139 <211> 11 <212> PRT <213><220><223> ^GFRLCDR1 <400> 139 Arg Ala Ser Gin Gly lie Asn Asn Tyr Leu Asn 1 5 10 ϋ > Λ > 01653 <21<21<21<21 140 7 PRT artificial sequence <220><223> Anti-EGFR LCDR2 <400> 140 Asn Thr Asn Asn Leu Gin Thr <210> 141 <211> 8 <212> PRT <213> Artificial Sequence <220><223> Anti-EGFRLCDR3 <400> 141 Leu Gin His Asn Ser Phe Pro Thr b > Λ > 012 3 1A n <2<2<2<2 142 120 PRT Artificial Sequence] 6156]-Order Table, doc Π8- 201247221 < 220 > < 223 > ^ EGFRVH < 400 > 142

Gin Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Thr Asp Tyr 20 25 30

Lys He His Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Tyr Phe Asn Pro Asn Ser Gly Tyr Ser Thr Tyr Ala Gin Lys Phe 50 55 60

Gin Gly Arg Val Thr lie Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Leu Ser Pro Gly Gly Tyr Tyr Val Met Asp Ala Trp Gly Gin 100 105 110

Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 143 <211> 108 <212> PRT <213> Artificial sequence <220><223> ^LEGFRVL <400>

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Arg Ala Ser Gin Gly lie Asn Asn Tyr 20 25 30

Leu Asn Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Arg Leu lie 35 40 45

Tyr Asn Thr Asn Asn Leu Gin Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gin His Asn Ser Phe Pro Thr 85 90 95

Phe Gly Gin Gly Thr Lys Leu Glu lie Lys Arg Thr 100 105 -119- 161561 - Sequence Listing.doc s 201247221 <210> 144 <211> 118 <212> PRT <213>Artificial Sequence<220><223> against "^-111\^(1) <400> 144

Gin Val Glu Leu Val Glu Ser Gly Gly Gly Val Val Gin Pro Gly Arg 15 10 15

Ser Gin Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Gly Met His Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ala He lie Trp Phe Asp Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95

Ala Arg Glu Leu Gly Arg Arg Tyr Phe Asp Leu Trp Gly Arg Gly Thr 100 105 110

Leu Val Ser Val Ser Ser 115 <210> 145 <21I> 108 <212> PRT <213>Artificial Sequence<220><223> Anti 10?-111\^(1) <400> 145

Glu He Val Leu Thr Gin Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie 35 40 45

Tyr Asp Ala Ser Lys Arg Ala Thr Gly lie Pro Ala Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Glu Pro 65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Arg Ser Lys Trp Pro Pro 85 90 95 -120- 161561 · Sequence Listing.doc 201247221

Trp Thr Phe Gly Gin Gly Thr Lys Val Glu Ser Lys 100 105 <210> 146 <211> 118 <212> PRT <213> Manual Sequence <220><223> Anti "^-111>/ 11(2) <400> 146

Gin Val Gin Leu Val Glu Ser Gly Gly Gly Val Val Gin Pro Gly Arg 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30

Gly Met His Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45

Ala lie lie Trp Phe Asp Gly Ser Ser Lys Tyr Tyr Gly Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Glu Leu Gly Arg Arg Tyr Phe Asp Leu Trp Gly Arg Gly Thr 100 105 110

Leu Val Thr Val Ser Ser 115 <210> 147 <211> 108 <212> PRT <213> Manual Sequence <220><223> Anti 10卩-111\^(2) <400> 147

Glu lie Val Leu Thr Gin Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 15 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie 35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly lie Pro Ala Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Glu Pro 65 70 75 80 .121 - 161561 · Sequence Listing.doc 201247221

Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Arg Ser Lys Trp Pro Pro 85 90 95

Trp Thr Phe Gly Gin Gly TTir Lys Val Glu lie Lys 100 105 >>> Λ > 01643 11 1i 1Λ 1i 148 5 PRT Artificial Sequence <220><223> Anti-CEAHCDR1 <400> 148 Glu Phe Gly Met Asn <210> 149 <211> 17 <212> PRT <213>Artificial Sequence<220><223> Anti-K £8 1 0112 <400> 149 Trp lie Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe Lys 15 10 15

Gly <210> 150 <211> 12 <212> PRT <213>Artificial sequence<220><223> Anti-CEAHCDR3 <400> 150 Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr 1 5 10 <210> 151 <211> 11 <212> PRT <213> artificial sequence <220><223><400> 151 Lys Ala Ser Gin Asn Val Gly Thr Asn Val Ala 1 5 10 ^ >>> 11 Tx n IX < 2 < 2 < 2 < 2 152 7 PRT Artificial Sequence 161561 · Sequence Listing. doc 122 - 201247221 <220><223> Anti-CEALCDR2 <400>

Ser Ala Ser Tyr Arg Tyr Ser <210> 153 <211> 10 <212> PRT <213>Artificial sequence <220><223><400>

His Gin Tyr Tyr Thr Tyr Pro Leu Phe Tlir 1 5 10 <210> 154 <211> 121 <212> PRT <213> Artificial Sequence <220><223> Anti-CEAVH <400>

Gin Val Gin Leu Val Gin Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala 15 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Phe 20 25 30

Gly Met Asn Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45

Gly Trp lie Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe 50 55 60

Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr 65 70 75 80

Leu Gin lie Ser Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Thr Val Thr Val Ser Ser 115 120 0>1>2>3>0>3>0> mil 2 o <2<2<2<2<2<2<4 155 108

PRT artificial sequence anti-CEAVL 155 123- 161561-sequence table.doc 201247221

Asp lie Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15

Asp Arg Val Thr lie Thr Cys Lys Ala Ser Gin Asn Val Gly Thr Asn 20 25 30

Val Ala Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu He 35 40 45

Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys His Gin Tyr Tyr Thr Tyr Pro Leu 85 90 95

Phe Thr Phe Gly Gin Gly Thr Lys Leu Glu lie Lys 100 105 0123 '*·_ -* · <2<2<2<2 68T person 1532 four wisdom <400> 156

Thr Lys GJy Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr 15 10 15

Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro 20 25 30

Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val 35 40 45

His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser 50 55 60

Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr He 65 70 75 80

Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Ala 85 90 95

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 100 105 110

Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 115 120 125

Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys Val Val 130 135 】 40

Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 145 150 155 160 -124- 161561 · Sequence Listing.doc 201247221

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gin 165 170 175

Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gin 180 185 190

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 195 200 205

Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys Gly Gin Pro 210 215 220

Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 225 230 235 240

Lys Asn Gin Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 245 250 255

Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn Tyr 260 265 270

Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 275 280 285

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val Phe 290 295 300

Ser Cys Ser Val Met His Glu Ala Leu His Ass His Tyr Thr Gin Lys 305 310 315 320

Ser Leu Ser Leu Ser Pro Gly Lys 325 .125- 161561-Sequence List.doc

Claims (1)

201247221 VII. Patent Application Range: 1. A combination of U) an immunoconjugate comprising at least one antigen-binding portion and an effector moiety and (b) an engineered antibody that increases effector function, the combination being used to treat an individual in need thereof The disease. 2. A combination of claim 1, wherein the effector moiety is a cytokine. 3_ A combination of claim 1 or 2, wherein the effector moiety is a cytokine selected from the group consisting of IL_2, GM-CSF, iFN-α, and IL-12. 4. A combination of claim 1 or 2, wherein the effect portion is IL_2. 5. The combination of claim 4, wherein the ratio _2 effect portion is a mutation ratio _2 effect portion 'which comprises at least one amino acid mutation, in particular, the amino acid substitution is compared to the unmutated IL-2 effect portion Reduce or eliminate this mutant IL-2 effect. The affinity for the α_subunit of the IL_2 receptor is retained but retains the affinity of the 1L-2 effector portion for the intermediate affinity to the _2 receptor. 6' is the combination of claim 1 or 2, wherein the antigen binding portion is an antibody or an antibody fragment. 7. The combination of claim 1 or 2, wherein the antigen binding moiety is selected from the group consisting of a Fab molecule and a scFv molecule. 8. Such as "monthly project!" Or a combination of 2, wherein the immunoconjugate comprises a first antigen binding portion and a second antigen binding portion. 9. The combination of the claim 8 and the second antigen-binding portion and the second antigen-binding portion are each a Fab molecule. 1〇.=The combination of the claim 8 'the effect portion is bound to the first antigen P knife>, the amino terminal peptide bond or the carboxy terminal peptide bond, and the second antigen binds the P knife and the 4 effect portion Or the first antigen-binding portion shares an amine-based peptide 161561.doc 201247221 bond or a carboxy-terminal peptide bond. 11. The combination of claim W2, wherein the immunoconjugate comprises an effector moiety (especially a single-stranded effector moiety) and a first Fab molecule and a second Fab molecule, wherein the effector moiety is bonded at its amine-terminated amino acid At the carboxy terminus of the heavy or light chain of the first Fab molecule, and wherein the effector moiety is attached to the amino terminus of the heavy or light bond of the second Fab molecule at its carboxyl guanamine group. 12 - A combination of a sputum or a combination of 2, wherein the antigen binding portion is directed against an antigen present on a tumor cell or in a tumor cell environment. 13. A combination of claim 1 or 2, wherein the engineered antibody that increases effector function is a full length IgG class antibody, particularly a "(1) subclass antibody. 14. A combination of claims or 2, wherein The increase in effector function is selected from the group consisting of an increase in binding to an activated Fc receptor, an increase in ADcc, an increase in adCP, an increase in CDC, and an increase in cytokine secretion. 1 5 . If a combination of claims 1 or 2, the effector function is increased to Increased binding to an activated Fc receptor and/or increased adcc. 16. The combination of claim 1 or 2, wherein the engineered to increase the effector function is introduced by introducing one or more amino acids in the Fc region Mutation or engineering by modification of glycosylation in the Fc region 17. As in the combination of claim 1 or 2, wherein the engineered antibody that increases the effector function b is engineered and unengineered The antibody increases the ratio of unfucosylated oligosaccharides in the Fc region compared to the antibody. 18. The combination of claim 1 or 2 wherein the engineered anti-system that increases the effector function is directed against the antigen present on the tumor cells. .doc 201247221 19_ A combination of claim 1 or 2, wherein the disease is a condition treatable by stimulating effector cell function, particularly cancer. 20. A combination of claim 1 or 2 wherein the individual is a mammal, especially a human. A pharmaceutical composition comprising (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) an engineered antibody that increases effector function in a pharmaceutically acceptable carrier 22. Use of (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) an antibody engineered to increase effector function for the manufacture of a medicament for treating a disease in an individual. 23. Use of (a) an immunoconjugate comprising at least one antigen binding portion and an effector moiety and (b) a combination of an engineered antibody that increases effector function for use in the manufacture of an effector cell function 24. A kit for treating a disease, the kit being contained in the same container or individual troughs (a) comprising at least one antigen binding portion and an effect portion An immunoconjugate, (b) an antibody engineered to increase effector function, and (c) a package insert as appropriate, the package insert containing printed instructions for directing the combination therapy as a method of treating the disease. Doc