SPECIFIC DOUBLE ANTIBODIES

Abstract

The invention provides dual-specific antibodies and methods for manufacturing and using these antibodies. In general, the dual-specific antibodies are generated by identifying a monospecific antibody with electrostatic or hydrophobic light chain variable region (VL) residues and altering the nucleic acid sequence encoding one or more solvent-accessible residues in the antibody's VH, either alone or in combination with altering the nucleic acid sequence encoding the antibody's VL. The altered VH and VL are expressed, and dual-specific antibodies or antigen-binding fragments are selected from them. Examples of dual-specific antibodies and methods for using the antibodies are also provided.

Description

SPECIFIC DOUBLE ANTIBODIES FIELD OF INVENTION The present invention relates to specific double antibodies and methods of manufacturing and using these antibodies. BACKGROUND OF THE INVENTION Antibodies are specific immunoglobulin polypeptides produced by the vertebrate immune system in response to stimulation by foreign proteins, glycoproteins, cells, or other foreign antigenic substances. A crucial part of this process is the generation of antibodies that bind specifically to a particular foreign substance. The binding specificity of these polypeptides to a particular antigen is highly refined, and the multitude of specificities that an individual vertebrate can generate is exceptional in its complexity and variability. Thousands of antigens can elicit responses, each directed almost exclusively to the specific antigen that triggered it. Specific antigen recognition is essential for antibody function in the adaptive immune response. The combinatorial association of a heavy chain (HC) and a light chain (LC) is conserved across all vertebrates in the generation of the antibody repertoire. However, there is an asymmetry of diversity between the two chains. The variable domain of the HC (Vh) contains considerably greater sequence diversity and contributes antigen recognition determinants more frequently than the variable domain of the LC (Vl). Nevertheless, given the variability with which antibodies recognize and bind to a particular foreign substance, some antibodies typically utilize Vl for antigen-binding energy. The specificity of antibodies and antibody fragments for one or more particular antigens makes them desirable therapeutic agents. Antibodies and antibody fragments can be used to target specific antigens, exhibiting pleiotropic biological functions (e.g., cytokines). Therefore, there is a current and ongoing need to identify and characterize therapeutic antibodies, especially antibodies, fragments, and their derivatives, useful in the treatment of various diseases and disorders, such as allergic diseases, inflammatory diseases, autoimmune diseases, and proliferative diseases. SUMMARY OF THE INVENTION The present invention provides methods for preparing specific double antibodies and antibody fragments. The invention also provides specific antibodies identified using these methods, as well as their use. RbCCnn / l 7Π7 / Β / Y In general, the methods of the invention involve diversifying the Vh of an antibody to generate specific double antibody variants that can be stably expressed in a gene library. In one embodiment, prior to antibody diversification, the antibody is characterized by a Vh and a Vl that pair to form an antigen-binding site that binds specifically to a first epitope, but not a second epitope. The antibody is further characterized by a hydrophobic or electrostatic residue on any one, two, or three of the amino acids located at positions 32, 50, or 91 (Kabat numbering system) of the Vl. This antibody with an electrostatic or hydrophobic residue at one or more of these positions is then altered at one or more amino acid residues (e.g., solvent-exposed amino acid residues) in the Vh. The Vh and Vl can then be expressed (e.g.,(like a gene library) and diversified specific double antibodies or antigen-binding fragments of these that can specifically bind to the first and second epitopes are selected from the expressed Vh and Vl. In one aspect, the invention presents a method for making a specific double antibody or antigen-binding fragment thereof, comprising a variable heavy chain domain (Vh) and a variable light chain domain (Vl), wherein the Vh and Vl of the specific double antibody are paired to form an antigen-binding site that binds specifically to a first epitope and a second epitope, wherein said method comprises the steps of: (a) providing an antibody comprising a Vh and Vl, wherein the Vh and Vl are paired to form an antigen-binding site that binds to a first epitope but not to the second epitope and wherein said antibody comprises at least one amino acid at position 32, 50 or 91 of the Vl that is electrostatic or hydrophobic, (b) altering the nucleic acid sequence encoding the Vh of the antibody of step (a), wherein one or more solvent-accessible amino acid residues are altered,(c) expressing the Vl and altered Vh of step (b) and (d) selecting a specific double antibody or antigen-binding fragment thereof, comprising the Vl and altered Vh of step (c), wherein the Vh and Vl are paired to form an antigen-binding site that binds specifically to the first epitope and the second epitope. In some embodiments, at least two of the amino acids at positions 32, 50, or 91 are either electrostatic or hydrophobic. In some embodiments, all three amino acids at positions 32, 50, and 91 are either electrostatic or hydrophobic. In some embodiments, the electrostatic residue is a tyrosine. In some embodiments, the hydrophobic residue is a tryptophan. In some embodiments, the nucleic acid sequence encoding Vh is altered according to the diversity of multiple naturally occurring heavy-chain amino acid sequences. In some embodiments, the solvent-exposed residue position is an amino acid residue position selected from the group consisting of positions 33, 34, 50, 58, and 95–97 of Vh. In some embodiments, the method further comprises altering the nucleic acid sequence encoding Vl of the antibody in step (a), where one or more solvent-accessible amino acid residues are altered.In some embodiments, the solvent-exposed residue position is a selected amino acid residue position from amino acids 93-96 of Vl. In some embodiments, altered Vhs are expressed in phages with Vl during the selection in step (d). In some embodiments, the antibody in step (a) comprises a light chain variable region complementarity-determining region CDRL 1 comprising the amino acid sequence KASQSVINDAA (SEQ ID NO: 9), a CDRL2 comprising the amino acid sequence YTSHRYT (SEQ. RbCCnn / l 7Π7 / Β / Y ID NO: 10) and a CDRL3 comprising the amino acid sequence QQDYTSPWTF (SEQ ID NO: 11). In some embodiments, the antibody of step (a) comprises a CDRH1 heavy chain variable region complementarity-determining region comprising the amino acid sequence DYSMH (SEQ ID NO: 13), a CDRH2 comprising the amino acid sequence VWINTETGEPTYADDFK (SEQ ID NO: 17), and a CDRH3 comprising the amino acid sequence GGIFYGMDY (SEQ ID NO: 20). In some embodiments, the antigen-binding site of the specific double antibody of step (d) binds to the first epitope and the second epitope mutually exclusively. In other embodiments, the antigen-binding site of the specific double antibody of step (d) binds to the first epitope and the second epitope simultaneously. In some forms, the first epitope comes from a biological molecule and the second epitope comes from the same biological molecule.In other embodiments, the first epitope is derived from a first biological molecule, and the second epitope is derived from a second biological molecule. In some embodiments, the first and second biological molecules are selected from the IL4 / IL5 and IL4 / IL13 groups. In some embodiments, the first and second biological molecules are cytokines. In some embodiments, the first or second biological molecule is a molecule that can increase the half-life of the dual-specific antibody when it binds to the antibody in vivo. In some embodiments, the first or second biological molecule is serum albumin or a neonatal iron receptor (FcRn). In some embodiments, the first or second biological molecule is a molecule that can enhance the effector function of a dual-specific antibody when it binds to the antibody in vivo.In some formulations, the first or second biological molecule binds to a cell surface protein on natural killer cells or macrophages. In some formulations, the cell surface protein is an Fe or C1q receptor. In some formulations, the Vh and Vl of the specific double antibody pair to form an antigen-binding site that binds specifically to the first or second epitope with a Kd of less than 10⁶. In some formulations, the Vh and Vl of the specific double antibody pair to form an antigen-binding site that binds specifically to the first or second epitope with a Kd of less than 10⁹. In some formulations, the Vh and Vl of the specific double antibody pair to form an antigen-binding site that binds specifically to the first or second epitope with a Kd of less than 10¹².In some modalities, the Vh and Vl of the specific double antibody pair to form an antigen-binding site that specifically binds to the first and second epitopes with a Kd of 10⁶⁰. In some modalities, the Vh and Vl of the specific double antibody pair to form an antigen-binding site that specifically binds to the first and second epitopes with a Kd of 10⁹⁰. In some modalities, the Vh and Vl of the specific double antibody pair to form an antigen-binding site that specifically binds to the first and second epitopes with a Kd of 10¹²⁰. In some modalities, the first and second biological molecules are not structurally similar. In some modalities, the selection of step (d) comprises deep sequencing, ultra-deep sequencing, and / or next-generation sequencing. Examples of antibodies produced using the methods of the invention include antibodies that bind to both interleukin 4 (IL4) and interleukin 5 (IL5), as well as antibodies that bind to both IL4 and interleukin 13 (IL13), RbCCnn / l 7P7 / B / Y as described below. The successful production of these antibodies demonstrates that altering the sequence of an antibody's heavy chain variable domain can serve as a general genetic modification pathway that generates antibodies with dual function and specificity. Dual-specific antibodies, including, but not limited to, the IL4 / IL5 and IL4 / IL13 antibodies described herein, have the potential to simultaneously target two pathways (redundant or non-redundant) and are useful for treating various diseases and disorders, including, but not limited to, immunological, inflammatory, and proliferative disorders. Accordingly, in another aspect, the invention presents an isolated specific double antibody or antigen-binding fragment thereof, prepared using the method of the invention described above. In some embodiments, the specific double antibody is a monoclonal antibody. In some embodiments, the fragment is a Fab or an scFv. In some embodiments, the specific double antibody is an IgG. In another aspect, the invention presents an isolated double specific antibody or antigen-binding fragment thereof comprising the amino acid sequence of any of the antibodies in Figures 4A, 4B, 4C, 7A, 7C, or 7D. In another aspect, the invention presents a specific double antibody isolated or antigen-binding fragment thereof, comprising the following six CDRs: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (iii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11), (iv) a CDRH1 comprising the amino acid sequence of DYDIH (SEQ ID NO: 14), (v) a CDRH2 comprising the amino acid sequence of VWINTETGEPTYADDFK (SEQ ID NO: 17), and (vi) a CDRH3 comprising the amino acid sequence of EILFYGMDY (SEQ ID NO: 21). In another aspect, the invention presents a specific double antibody isolated or antigen-binding fragment thereof, comprising the following six CDRs: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (iii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11), (iv) a CDRH1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15), (v) a CDRH2 comprising the amino acid sequence of AGIVYDATGFTTYADDFK (SEQ ID NO: 18), and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20). In another aspect, the invention presents an isolated double specific antibody or antigen-binding fragment thereof, comprising the following six CDRs: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (iii) a CDRL3 comprising the amino acid sequence of QQDYTPFPLTF (SEQ ID NO: 12), (iv) a CDRH1 RbCCnn / l 7Π7 / Β / Y comprising the amino acid sequence of DYLMH (SEQ ID NO: 16), (v) a CDRH2 comprising the amino acid sequence of AVIVSITGRTYYADDFK (SEQ ID NO: 19) and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20). In another aspect, the invention presents a specific double antibody isolated or antigen-binding fragment thereof, comprising the following six CDRs: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (iii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11), (iv) a CDRH1 comprising the amino acid sequence of DYSMH (SEQ ID NO: 13), (v) a CDRH2 comprising the amino acid sequence of GVIFQSGATYYADDFK (SEQ ID NO: 22), and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20). In another aspect, the invention presents a specific double antibody isolated or antigen-binding fragment thereof, comprising the following six CDRs: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (iii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11), (iv) a CDRH1 comprising the amino acid sequence of DYSMH (SEQ ID NO: 13), (v) a CDRH2 comprising the amino acid sequence of GIIFYTGHTYYADDFK (SEQ ID NO: 23), and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20). In another aspect, the invention presents an isolated double specific antibody or antigen-binding fragment thereof, comprising the following six CDRs: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (iii) a CDRL3 comprising the amino acid sequence of QQDYX1X2PWTF (SEQ ID NO: 24), where X1 is Thr, Ie, Leu or Lys and X2 is Ser or His; (iv) a CDRL1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15); (v) a CDRH2 comprising the amino acid sequence X1GIVYDATGFTX2YA X3X4FK (SEQ ID NO: 25), where X1 is Ala or Gly, X2 is Thr, Lie, Val or Ala, X3 is Asp, Val or Glu, and X4 is Asp, Glu, Asn, Ser, Lie, Leu, Thr, Ala or Phe; and (vi) a CDRH3 comprising the amino acid sequence GGIFYGMDY (SEQ ID NO: 20).In some embodiments, the specific double antibody or antigen-binding fragment thereof comprises the following six CDRs: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (iii) a CDRL3 comprising the amino acid sequence of QQDYTHPWTF (SEQ ID NO: 27), (iv) a CDRH1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15), (v) a CDRH2 comprising the amino acid sequence of GGIVYDATGFTTYAEEFK (SEQ ID NO: 28), and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20). In some embodiments, the specific double antibody or antigen-binding fragment thereof comprises the following six CDRs: (i) a. Rbccnn / ι ζηζ / E / γ CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9), (i) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10), (ii) a CDRL3 comprising the amino acid sequence of QQDYKHPWTF (SEQ ID NO: 31), (iv) a CDRH1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15), (v) a CDRH2 comprising the amino acid sequence of AGIVYDATGFTVYADDFK (SEQ ID NO: 32) and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20). In some forms, the specific double antibody or antigen-binding fragment thereof further comprises a frame region 3 (FR3) comprising the amino acid sequence GRX1TITX2DX3STSTX4 (SEQ ID NO: 26), where X1 is Val or Phe, X2 is Arg or lie, X3 is Thr, Phe, Met or Pro, and X4 is Ala or Val. In another aspect, the invention presents a specific double antibody or antigen-binding fragment thereof comprising a light-chain variable region selected from the amino acid sequences SEQ IDs 1, 5, 29, or 33 and a heavy-chain variable region selected from SEQ IDs 2, 3, 4, 6, 7, 8, 30, or 34. In some embodiments, the antibody or antigen-binding fragment thereof binds to IL4 with a Kd of 500 nM or less and to IL5 with a Kd of approximately 900 nM or less. In some embodiments, the antibody or antigen-binding fragment thereof binds to IL4 with a Kd of 100 nM or less and to IL5 with a Kd of approximately 100 nM or less. In some modalities, the antibody or antigen-binding fragment of it binds to IL4 with a Kd of 10 nM or less and to IL5 with a Kd of around 50 nM or less.In other formulations, the isolated double-specific antibody or antigen-binding fragment binds to IL4 with a Kd of 500 nM or less and to IL13 with a Kd of approximately 900 nM or less. In some formulations, the antibody or antigen-binding fragment binds to IL4 with a Kd of 100 nM or less and to IL13 with a Kd of approximately 100 nM or less. In some formulations, the antibody or antigen-binding fragment inhibits or blocks the binding of IL4, IL5, or IL13 to its receptor. In some formulations, the antibody is a monoclonal antibody. In certain formulations, the antibody is an IgG antibody. In some formulations, the antigen-binding fragment is a Fab fragment or a single-chain variable fragment (scFv). In some formulations, at least part of the frame sequence is a human consensus frame sequence. In some forms, the antibody is a chimeric, humanized, or fully human antibody. A pharmaceutical composition comprising any of the above specific double antibodies or antigen-binding fragments thereof is also provided. In another aspect, the invention presents an isolated nucleic acid encoding any of the specific double antibodies described herein, comprising a vector (e.g., an expression vector) for expressing the antibody. In another aspect, the invention features host cells comprising the vectors and / or nucleic acids described above. In some embodiments, the host cell is a mammalian cell (e.g., a Chinese hamster ovary (CHO) cell). In other embodiments, the host cell is a prokaryotic cell (e.g., an E. coli cell). A method for producing any of the specific double antibodies is also provided. Rbccnn / ι znz / R / v above, where the method comprises culturing the host cell that produces the double specific antibody and recovering the double specific antibody from the host cell or the culture medium. In another aspect, the invention presents a method for treating asthma in a subject, wherein the method comprises administering to the subject any of the specific double antibodies described herein, wherein the administration is carried out for a time and in a quantity sufficient to treat or prevent asthma in the subject. In some embodiments, the method also comprises administering at least one additional asthma treatment selected from the group consisting of an IgE antagonist, an antihistamine, threophylline, salbutamol, beclomethasone dipropionate, sodium cromoglycate, a steroid, and anti-inflammatory agents. In some embodiments, the asthma is allergic asthma. In yet another aspect, the invention presents a method for treating a proliferative disorder in a subject, wherein the method comprises administering to the subject any of the specific double antibodies described herein, wherein the administration is carried out for a time and in a quantity sufficient to treat the proliferative disorder in the subject. In some embodiments, the proliferative disorder is cancer. In some embodiments, the method also comprises administering to the subject an additional antiproliferative agent selected from the group consisting of a chemotherapeutic agent, a cytotoxic agent, and an antiangiogenic agent. BRIEF DESCRIPTION OF THE DRAWINGS Figures 1A and 1B are graphs showing the CDR mutagenesis correspondence of the hu19C11 antibody. To measure the relative antigen-binding affinity of Fab variants of hu19C11, the binding of alanine mutans of LC CDR or wild-type (wt) hu19C11 anti-IL4 expressed serially diluted phages to anti-gD (A) or IL4 (B) antibody coated in ELISA wells was detected by Fab expression and a horseradish peroxidase conjugate (HRP) on anti-M phages to quantify antigen binding, respectively. gD is a peptide tag fused to the C-terminus of the light chain. The anti-gD antibody was coated directly onto the ELISA wells, while IL4 was captured with an unblocked anti-IL4 antibody coated onto ELISA wells. Figure 1C is a graph showing that the effects of alanine mutations at individual CDR sites were examined by comparing the relative IL4 binding affinity of Fab expression variants in phages using analyses similar to those in Figures 1A and 1B. Single-letter coded amino acids were used. Relative IL4 binding resistances were determined by fitting the data to a linear regression model and then dividing the IL4 binding slope (versus phage concentration) by the Fab expression slope. Low values ​​(below the dashed line) were considered to indicate low IL4 binding affinity relative to hu 1911 wt, suggesting disruptive mutations. Figure 2A is a structural diagram showing the correspondence of critical IL4 binding residues of hu19C11 in a top-down view of the Fab structure trastuzumab (PDB: 1FDV). The model The structural RbCCnn / l 7P7 / B / Y of hu19C11 was generated using MOE with input information from POB 3SQO and 3BEI as templates for the heavy and light chains, respectively. The residues important for IL4 binding are LC residues 31, 32, 50, 53, 91, and 92, and HC residues 31, 32, 96, 98, and 99 (in Kabat numbering). These residues are shown in red in the model structure, viewed vertically from the top of the antigen-binding site. Libraries of Fab hu19C11 variants expressed in phages were generated by site-directed mutation. HC residues that were allowed the 20 amino acids preferentially for wild-type residues are labeled in bold, while other HC and LC residues, depending on the label, were allowed limited mutation to mimic natural diversity. Figure 2B is a table showing the design of combination libraries for recruiting secondary antigen specificity for hu19C11. CDR sequences from selected clones are shown, including specific anti-IL5 (5A), specific anti-IL4 / 5 (E7, B1), and specific anti-IL4 / 13 (F1, F2) antibody clones with mutations from the original wild-type hu19C11. Randomly separated residues in the libraries and mutations in the isolated clones are shaded according to their properties: Y, W, F aromatic side chain; I, V, A, M, L hydrophobic; R, H, K basic; E, D acidic; T, N, Q, S polar; and P, G. Relative antigen-binding affinity is shown as measured by IC50 from phage competence assays.The Fab-expressing phage was first incubated with serial dilutions of the respective antigens in solution for 2 h. The unbound phage was then briefly captured using antigen-coated ELISA wells and detected with anti-M13 HRP conjugate. The antigen concentration that inhibits 50% of phage binding to the antigen-coated well is calculated as IC50. NB denotes the absence of detectable binding of phage clones directly to the antigen-coated wells. Figure 3 is a graph showing a titration of ten gene libraries generated by expression in phages (2144-1 to 2144-10) for IL4 binding capacity. Figure 4A shows the light chain variable domain amino acid sequence alignment of double-specific anti-IL4 / IL5 variants of hu19C11 (E7 (SEQ ID NO: 1) and B1 (SEQ ID NO: 5)), as well as 5A specific anti-IL5 (SEQ ID NO: 1), aligned with respect to hu19C11 specific anti-IL4 (SEQ ID NO: 1). Figure 4B shows the heavy chain variable domain amino acid sequence alignment of double-specific anti-IL4 / IL5 variants of hu 19C11 (E7 (SEQ ID NO: 4) and B1 (SEQ ID NO: 6)), as well as 5A specific anti-IL5 (SEQ ID NO: 3), aligned with respect to hu19C11 specific anti-IL4 (SEQ ID NO: 2). Figure 4C shows the heavy chain variable domain amino acid sequence alignment of double-specific anti-IL4 / IL13 variants of hu19C11 (F1 (SEQ ID NO: 7) and F2 (SEQ ID NO: 8)), aligned with respect to anti-IL4 specific hu19C11 (SEQ ID NO: 2). RbCCnn / l 7Π7 / Β / Y Figure 5 is a graph showing the dual specificity of selected hu19C11 variants as IgG. The antigen-binding specificity of selected hu19C11 variants was assessed as the binding of these variants in the form of 250 nM human IgG1 to one or more target antigens or various irrelevant coated proteins in ELISA wells with detection by anti-Fc antibody HRP conjugates. Figure 6 is a graph showing the binding specificity characterization of specific double variants of hu19C11 (wild-type specific anti-IL4: 19C11; specific anti-IL5: 5A1; and specific anti-IL4 / 5: E7 and B1) at the indicated antibody concentrations in the blocking interaction of IL5 with its receptor. The levels of biotinylated IL5 receptor bound to IL5 immobilized on ELISA wells in the presence of increasing concentrations of humanized 19C11 or variants such as IgG were detected with HRP-streptavidin conjugate. Figure 7A is a table showing the amino acid sequence of mature affinity variants of IL4 / IL5-specific E7 and their relative affinity to target antigens as measured by phage competence assays. To improve affinity, phage libraries expressing E7 variants were constructed using selected mutated residues with the random separation strategies "homologous" (bold), "limited" (italicized), and "soft" (gray), respectively. These strategies allow for wild-type and homologous amino acids, limited diversity based on natural antibodies, or approximately 50% wild-type and 50% of all other amino acids. CDR H2 was primarily found in homologous mutation to subtly optimize the newly recruited binding function with respect to IL5. For other CDRs, the target was non-critical sites for IL4 binding. The sequences of selected clones were aligned with respect to E7, and mutations are shown.The relative affinity of each clone was assessed using the IC50 of phages as indicated above. Figure 7B is a table showing the affinities of E7 and its affinity-matured variants, 1C36 and 1C60, purified as Fabs, as measured by Biacore using a CM5 sensor chip immobilized with human IL5 (R&D Systems) or IL4 at 25C. Figure 7C shows the light chain variable domain amino acid sequence alignment of affinity-enhancing double anti-IL4 / IL5 specific variants of E7, 1C36 (SEQ ID NO: 29) and 1C60 (SEQ ID NO: 33), aligned with respect to the light chain variable domain amino acid sequence of E7 (SEQ ID NO: 1). Figure 7D shows the heavy chain variable domain amino acid sequence alignment of affinity-enhancing double anti-IL4 / IL5 specific variants of E7, 1C36 (SEQ ID NO: 30) and 1C60 (SEQ ID NO: 34), aligned with respect to the heavy chain variable domain amino acid sequence of E7 (SEQ ID NO: 4). Figures 8A and 8B are graphs showing the binding specificity characterization of E7 and E7 affinity-enhancing variants (1C36 and 1C60). Direct binding of E7 and E7 affinity-enhancing variants such as Rbccnn / ι ζηζ / E / γ IgG (100 nM) with respect to immobilized antigen and irrelevant proteins on an ELISA plate was detected by anti-IgG-HRP(A). The binding of biotinylated IL5 to the coated IL5 receptor in ELISA wells in the presence of buffer (PBS) or 50 nM of E7,1C36 or 1C60 was detected by streptavidin-HRP conjugate(B). DETAILED DESCRIPTION OF THE INVENTION Many disease pathways evolve through the action of more than one protein or a protein with more than one function. For example, allergic, inflammatory, or autoimmune disorders (e.g., asthma) often involve multiple cytokines. Dual-specific antibodies are useful in both diagnostic and therapeutic applications where targeting more than one antigen is desired. The inventors discovered a novel method for generating dual-specific antibodies. When the antibody's Vl includes residues essential for antibody-antigen interaction, these antibodies can then be diversified by altering the Vh residues alone or in combination with additional framework and Vl residues. In general, the methods of the invention involve diversifying the Vh of an antibody to generate variants that can be stably expressed in a gene library. Specifically, an antibody that binds specifically to a first epitope but not to a second epitope and that is characterized by an electrostatic or hydrophobic residue on any one, two, or three of the amino acids at positions 32, 50, or 91 (in Kabat numbering) of the Vl is altered in one or more amino acid residues (e.g., solvent-exposed amino acid residues) in the Vh. The Vh and Vl are then expressed, and the diversified specific double antibodies or antigen-binding fragments thereof that can bind specifically to the first and second epitopes are then selected from the expressed Vh and Vl. Examples of antibodies produced using the methods of the invention include antibodies that bind to both interleukin 4 (IL4) and interleukin 5 (IL5), as well as antibodies that bind to both IL4 and interleukin 13 (IL13), as described below. These antibodies demonstrate that mutations in the heavy chain variable domain (e.g., CDRs) of an IL4 antibody confer dual-binding capabilities for additional unrelated proteins, as well as IL4, and provide proof of concept for the general strategy of altering residues in the Vh to confer dual specificity.Specific double antibodies, including, but not limited to, IL4 / IL5 and IL4 / IL13 antibodies described herein, have the potential to simultaneously target multiple antigens, e.g., redundant and non-redundant cytokine pathways, making them useful for the treatment of various diseases and disorders, including cytokine-mediated diseases (e.g., asthma). I. Definitions The term "multispecific antibody" is used in the broadest sense and specifically covers an antibody comprising a heavy chain variable domain (Vh) and a light chain variable domain (Vl), where the VhVl unit has polyepitopic specificity (i.e., it can bind to two different epitopes on a biological molecule or to each epitope). RbCCnn / l 7P7 / B / Y in a different biological molecule). These multispecific antibodies include, but are not limited to, full-length antibodies, antibodies with two or more Vl and Vh domains, antibody fragments such as Fab, Fv, dsFv, scFv, diabodies, bispecific diabodies, and tribodies, and covalently or non-covalently linked antibody fragments. “Polyepitope specificity” refers to the ability to bind specifically to two or more different epitopes on one or more identical or different targets. “Dual specificity” or “bispecificity” refers to the ability to bind specifically to two different epitopes on one or more identical or different targets. However, unlike bispecific antibodies, double specific antibodies are naturally occurring IgG antibodies in which the two antigen-binding groups are identical in amino acid sequence, and each Fab group can recognize two antigens.Dual specificity allows antibodies to interact with high affinity with two different antigens as a single Fab or IgG molecule. According to one model, the multispecific antibody in an IgG1 form binds to each epitope with an affinity of 5 μM to 0.001 pM, 3 μM to 0.001 pM, 1 μM to 0.001 pM, 0.5 μM to 0.001 pM, or 0.1 μM to 0.001 pM. "Monospecific" refers to the ability to bind to only one epitope. In general, an antibody includes a 4-chain basic antibody unit, which is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. (An IgM antibody consists of 5 of the basic heterotetrameric units along with an additional polypeptide called the J chain and thus contains 10 antigen-binding sites, while secreted IgA antibodies can polymerize to form polyvalent assemblies comprising 2–5 of the 4-chain basic units along with the J chain.) In the case of IgG, the 4-chain unit is typically around 150,000 daltons. Each L chain is linked to an H chain by a covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds, depending on the H chain isotype. Each H and L chain also has intrachain disulfide bridges separated at regular intervals.Each H chain has a variable domain (Vh) at its N-terminus, followed by three constant domains (Ch) for each of the oyyy chains and four CH domains for the μ and ε isotypes. Each L chain has a variable domain (Vl) at its N-terminus, followed by a constant domain (Cl) at its other end. The Vl is aligned with the Vh, and the Cl is aligned with the first constant domain of the heavy chain (Ch1). Particular amino acid residues are thought to form an interface between the variable domains of the light and heavy chains. The pairing of a Vh and Vl forms an antigen-binding site. For the structure and properties of the different classes of antibodies, refer, e.g., to Basic and Clinical Immunology, 8th edition, Daniel P. Stites, Abba I. Terry, Tristram G. Parslow (eds.), Appleton & Lange, Norwalk, CT, 1994, page 71 and chapter 6.The L chain of any vertebrate species can be assigned to one of two distinct types, designated kappa and lambda, based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains (Ch), immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, with heavy chains designated α, δ, ε, γ, and μ, respectively. The γ classes are further divided into subclasses based on relative differences. Rbccnn / ι ζηζ / E / γ minors in the sequence and function of Ch, e.g., humans express the following subclasses: lgG1, lgG2, lgG3, lgG4, lgA1 and lgA2. The term “variable” refers to the fact that certain segments of the variable domains differ widely in sequence among antibodies. The variable domain, or “V,” mediates antigen binding and defines the specificity of a particular antibody for its particular antigen. However, this variability is not uniformly distributed across a 110-amino-acid span of the variable domains. Instead, V regions consist of relatively invariable spans called frame regions (FRs) of 15–30 amino acids separated by shorter regions of extreme variability called “hypervariable regions,” each 9–12 amino acids long.Each of the variable domains of natural heavy and light chains comprises four receptors (RFs), which primarily adopt a beta-sheet configuration, connected by three hypervariable regions. These hypervariable regions form loops that connect, and in some cases integrate, the beta-sheet structure. The hypervariable regions in each chain are held close together by the RFs and, with the hypervariable regions of the other chain, contribute to the formation of the antigen-binding site of antibodies (refer to Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991)). The constant domains are not directly involved in antibody-antigen binding but have various effector functions, such as antibody participation in antibody-dependent cellular cytotoxicity (ADCC). The term "hypervariable region", when used herein, refers to amino acid residues of an antibody that are responsible for antigen binding. The hypervariable region generally comprises amino acid residues from a "complementarity-determining region" or "CDR" (e.g., around approximately residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in V1 and around approximately residues 26-35 (H1), 49-65 (H2), and 95-102 (H3) in V2 (in one modality, H1 is found around approximately residues 31-35); Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and / or residues forming a "hypervariable loop" (e.g., residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in V1 and 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the VH; Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). The term “monoclonal antibody,” as used herein, refers to an antibody from a substantially homogeneous antibody population; that is, the individual antibodies comprising the population are substantially similar and bind to the same epitope or epitopes, except for possible variants that may arise during the production of the monoclonal antibody, with these variants generally present in minor quantities. This monoclonal antibody typically includes an antibody comprising a variable region that binds to a target, the antibody being obtained by a process that includes the selection of the antibody from among multiple antibodies. For example, the selection process may be the selection of a single clone from among multiple clones, such as a set of hybridoma clones, phage clones, or recombinant DNA clones. It should be understood that the The selected RbCCnn / l 7P7 / B / Y antibody may be further altered, for example, to improve its affinity for the target, to humanize the antibody, to improve its production in cell culture, to reduce its immunogenicity in vivo, to create a multispecific antibody, etc., and an antibody comprising the altered variable region sequence is also a monoclonal antibody of the present invention. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are normally not contaminated by other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody obtained from a substantially homogeneous antibody population and should not be interpreted as requiring the production of the antibody by any particular method.For example, the monoclonal antibodies for use according to the present invention can be made using various techniques including the hybridoma method (e.g., Kohler et al., Nature, 256:495 (1975); Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681, (Elsevier, NY, 1981), recombinant DNA methods (refer, e.g., to U.S. Patent No. 4,816,567), phage expression technologies (refer, e.g., to Clackson et al., Nature, 352:624-628 (1991); Marks et al., J. Mol. Biol., 222:581-597) (1991); Sidhu et al, J. Mol. Biol. 338(2):299-310 (2004); Lee et al, J.Mol.Biol.340(5): 1073-1093 (2004); Fellouse, Proc. Nat. Acad. Sci. USA 101(34):12467-12472 (2004); and Lee et al. J. Immunol.Methods 284(1-2):119-132 (2004) and technologies for producing human or human-like antibodies from animals with parts or all of the human immunoglobulin genes or loci encoding human immunoglobulin sequences (refer, e.g., to WO98 / 24893, WO / 9634096, WO / 9633735 and WO / 91 10741, Jakobovits et al., Proc. Nati. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immuno, 7:33 (1993); U.S. patent nos. 5,545,806, 5,569,825, 5,591,669 (all from GenPharm); 5,545,807; WO 97 / 17852, US Patent No. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016, and Marks et al, Bio / Technology, 10:779-783 (1992); Lonberg et al, Nature, 368: 856-859 (1994); Morrison, Nature, 368: 812-813 (1994); Fishwild et al, Nature Biotechnology, 14: 845-851 (1996); Neuberger, Nature Biotechnology, 14: 826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol, 13: 65-93 (1995). Monoclonal antibodies herein specifically include chimeric, humanized, fully human, and affinity-maturated antibodies. Chimeric antibodies are antibodies in which a portion of the heavy and / or light chain is identical or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, provided they exhibit the desired biological activity (U.S. Patent No. 4,816,567; and Morrison et al., Proc. Nati. Acad. Sci. USA, 81:6851-6855 (1984)).The chimeric antibodies of interest herein include "primatized" antibodies comprising variable domain antigen-binding sequences derived from a non-human primate (e.g., old world monkey, ape, etc.) and human constant region sequences. RbCCnn / l 7Π7 / Β / Y Humanized forms of non-human antibodies (e.g., rodent) are chimeric antibodies containing a minimal amount of sequence derived from the non-human antibody. Mostly, humanized antibodies are human immunoglobulins (the recipient antibody) in which residues from a hypervariable region of the recipient are replaced with residues from a hypervariable region of a non-human species (the donor antibody), such as mouse, rat, rabbit, or non-human primate, with the desired specificity, affinity, and capacity. In some cases, residues from the framework region (FR) of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, humanized antibodies may include residues not found in either the recipient or donor antibody. These modifications are made to further refine the antibody's performance.In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, where all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the receptors are those of a human immunoglobulin sequence. The humanized antibody will optionally also comprise at least a portion of an immunoglobulin constant region (Fe), typically that of a human immunoglobulin. For further details, refer to Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). A "human antibody" is one that possesses an amino acid sequence corresponding to that of an antibody produced by a human and / or made using techniques for manufacturing human antibodies. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. A "matured affinity" antibody is one with one or more alterations in one or more of its CDRs that produce an improvement in the antibody's affinity for the antigen, compared to an original antibody without such alteration(s). Preferred matured affinity antibodies will have nanomolar or even picomolar affinities for the target antigen. Matured affinity antibodies are produced by procedures known in the art. Marks et al. Bio / Technology 10:779-83 (1992) describe affinity maturation by transposition of Vh and Vl domains. Random mutagenesis of CDRs and / or frame residues is described in: Barbas et al., Proc Nat. Acad. Sel. USA 91:3809-13 (1994); Schier et al. Gene 169:147-55 (1995); Yelton et al., J. Immunol. 155:1994-2004 (1995). Jackson et al., J. Immunol. 154(7):3310-19 (1995); and Hawkins et al., J. Mol. Biol. 226:889-96 (1992). An "intact" antibody is one that comprises an antigen-binding site, as well as a Cl and at least the heavy-chain constant domains Ch1, Ch2, and Ch3. The constant domains may be constant domains of a natural sequence (e.g., constant domains of a human natural sequence) or an amino acid sequence variant thereof. Preferably, the intact antibody has one or more effector functions. RbCCnn / l 7Π7 / Β / Y “Antibody fragments” comprise a portion of an intact antibody, preferably the variable or antigen-binding region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (see U.S. Patent No. 5,641,870, Example 2; Zapata et al., Protein Eng. 8(10): 1057-1062 (1995)); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. The term "linear antibodies" generally refers to the antibodies described in Zapata et al., Protein Eng., 8(10):1057-1062 (1995). In summary, these antibodies comprise a pair of double Fd segments (Vh-Ch1-Vh-Ch1) which, together with complementary light chain polypeptides, form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific. Papain digestion of antibodies yields two identical antigen-binding fragments, termed Fab fragments, and a residual Fe fragment, named for their ability to readily crystallize. The Fab fragment consists of a complete L chain along with the variable region domain of the H chain (Vh) and the first constant domain of a heavy chain (Ch1). Treatment of an antibody with pepsin produces a single large F(ab j2) fragment that roughly corresponds to two disulfide-linked Fab fragments with bivalent antigen-binding activity and retains the ability to crosslink antigens. Fab' fragments differ from Fab fragments in that they have a few additional residues at the carboxy terminus of the Ch1 domain, which includes one or more cysteines from the antibody's hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine ​​residues of the constant domains bear a free thiol group.The F(abj2) antibody fragments were originally produced as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known. The Fe fragment comprises the carboxy-end portions of both H chains held together by disulfide bonds. The effector functions of antibodies are determined by the sequences in the Fe region, which is also the region recognized by Fe receptors (FCRs) found on certain cell types. The "Fv" domain consists of a dimer of a heavy-chain variable region domain and a light-chain variable region domain in close, non-covalent association. Six hypervariable loops (three loops each from the heavy and light chains) arise from the folding of these two domains. These loops contribute amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv domain with only three antigen-specific variable regions) can recognize and bind to the antigen, albeit at a lower affinity than the entire binding site. Single-chain Fvs, also abbreviated as sFv or scFv, are antibody fragments comprising the Vh and Vl antibody domains linked in a single polypeptide chain. Preferably, sFv polypeptides further comprise a polypeptide linker between the Vh and Vl domains, enabling the sFv to form the structure RbCCnn / l 7P7 / B / Y desired for antigen binding. For an sFv analysis, refer to Pluckthun in The Pharmacology of Monoclonal Antibodies, Volume 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994); Borrebaeck 1995. The term “diabodies” refers to small antibody fragments prepared by constructing sFv fragments (see previous paragraph) with short linkers (of about 5–10 residues) between the Vh and Vl domains to achieve inter-chain pairing but not inter-chain pairing of the V domains, resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. Bispecific diabodies are heterodimers of two “crossover” sFv fragments in which the Vh and Vl domains of the two antibodies are present on different polypeptide chains. Diabodies are described in more detail, for example, in EP 404,097; WO 93 / 11161; and Hollinger et al., Proc. Nati. Acad. Sel. USA, 90:6444–6448 (1993). "Electrostatic" means charged. Generally, electrostatic amino acids have charged or polar side chains. Examples of amino acids with polar side chains include serine, threonine, tyrosine, cysteine, asparagine, and glutamine. Examples of amino acids with negatively charged side chains include aspartic acid and glutamic acid. Examples of amino acids with positively charged side chains include lysine, arginine, and histidine. "Hydrophobic" means incompatible with water or that it does not readily dissolve, absorb, or mix in water. Generally, hydrophobic amino acids have nonpolar side chains, and examples include alanine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine. The term "cytokine" is a generic term for proteins released by one cell population that act on another cell as intercellular mediators. Examples of cytokines include lymphokines, monokines, and traditional polypeptide hormones. Cytokines include growth hormones, such as human growth hormone, human N-methionyl growth hormone, and bovine growth hormone; parathyroid hormone; thyroxine; insulin and proinsulin; relaxin and prorelaxin; glycoprotein hormones such as follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), and luteinizing hormone (LH); liver growth factor; fibroblast growth factor; prolactin; placental lactogen; tumor necrosis factor-α and -β; Müllerian inhibitory substance; and mouse gonadotropin-associated peptide. inhibin; activin;vascular endothelial growth factor; integrin; thrombopoietin (TRO); nerve growth factors such as NGF-β; platelet growth factor; transforming growth factors (TGFs), such as TGF-α and TGF-β; insulin-like growth factor I and II; erythropoietin (EPO); osteoinductive factors; interferons such as interferon-α, -β, and -γ; colony-stimulating factors (CSFs), such as macrophage-CSF (M-CSF); granulocyte-macrophage-CSF (GM-CSF); and granulocyte-CSF (G-CSF); interleukins (IL) such as IL-11, IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-11, IL-12, and IL-13; a tumor necrosis factor such as TNF-α or TNF-β; Rbccnn / ι znz / R / Y and other polypeptide factors including LIF and the kit ligand (KL). As used herein, the term cytokine includes proteins of natural or recombinant cell culture origins and biologically active equivalents of naturally occurring sequence cytokines. As used herein, "codon set" refers to a set of different triple nucleotide sequences used to encode desired variant amino acids. A set of oligonucleotides can be synthesized, for example, by solid-phase synthesis, including sequences representing all possible combinations of triple nucleotide structures provided by the codon set and encoding the desired group of amino acids. A standard form of codon designation is the IUB code, which is known in the art and described herein. A codon set is typically represented by three italicized capital letters, e.g., NNK, NNS, XYZ, DVK, and the like (e.g., the codon NNK refers to N-NTIGIC at positions 1 and 2 in the codon and K = G / T in an equimolar ratio at position 3 to encode the 20 naturally occurring amino acids).A “non-random codon set,” as used herein, therefore refers to a set of codons encoding selected amino acids that partially, preferably, or completely meet the criteria for amino acid selection as described herein. The synthesis of oligonucleotides with “degeneracy” of selected nucleotides at specific positions is known in the art, for example, the TRIM approach (Knappek et al., J. Mol. Biol. 296:57-86, 1999; Garrard and Henner, Gene 128:103, 1993). These oligonucleotide sets with specific codon sets can be synthesized using commercial nucleic acid synthesizers (available, for example, from Applied Biosystems, Foster City, CA) or can be obtained commercially (for example, from Life Technologies, Rockville, MD).Therefore, a set of oligonucleotides synthesized with a particular set of codons typically includes multiple oligonucleotides with different sequences, the differences being established by the codon set within the overall sequence. The oligonucleotides, as used according to the invention, have sequences that allow hybridization with a variable-domain nucleic acid template and may also, although not necessarily, include enzymatic restriction sites useful, for example, for cloning purposes. An antibody that "binds to" an antigen of interest is one that binds to the antigen with sufficient affinity to make the antibody useful as a diagnostic and / or therapeutic agent for targeting a protein, cell, or tissue that expresses the antigen and does not exhibit significant cross-reactivity with other proteins. In such cases, the extent of antibody binding to a "non-target" protein will be less than approximately 10% of the antibody binding to its specific target protein, as determined by fluorescence-activated cell sorting (FACS), radioimmunoprecipitation (RIA), or ELISA. Regarding antibody binding to a target molecule, the terms "specific binding," "specifically binds to," or "specific to" a particular polypeptide or epitope or a particular polypeptide target indicate a binding that is significantly different from a non-specific interaction.Specific binding can be measured, for example, by determining the binding of a molecule compared to. RbCCnn / l 7P7 / B / Y the binding of a control molecule. For example, specific binding can be determined by competition with a control molecule similar to the target, e.g., an excess of unlabeled target. In this case, specific binding is indicated if the binding of the labeled target to a probe is competitively inhibited by the excess of unlabeled target.The term "specific binding" or "specific binding" or "specific to" a particular polypeptide or epitope on a particular polypeptide target, as used herein, may be presented, for example, in a molecule with a target Kd of 10⁴M or less, alternatively 10⁵M or less, alternatively 10⁶M or less, alternatively 10⁷M or less, alternatively 10⁸M or less, alternatively 10⁹M or less, alternatively 10¹⁰M or less, alternatively 10¹¹M or less, alternatively 10¹²M or less, or a Kd ranging from 10⁴M to 10¹²M, or 10⁻⁶M to 10⁻¹⁰M, or 10⁻⁷M to 10⁻⁹M. As those skilled in the art will observe, affinity and Kd values ​​are inversely related. A high affinity for an antigen is measured by a low Kd value.In one embodiment, the term "specific binding" refers to binding in which a molecule binds to a particular polypeptide or epitope in a particular polypeptide without substantially binding to any other polypeptide or epitope of the polypeptide. "Biologically active" and "biological activity", as well as "biological characteristics", with respect to a polypeptide of the present invention, mean that it has the ability to bind to a biological molecule, except where otherwise specified. "Biological molecule" refers to a nucleic acid, a protein, a carbohydrate, a lipid, and combinations thereof. In one sense, the biological molecule exists in nature. "Isolate," when used to describe the various antibodies described herein, means an antibody identified and separated and / or recovered from a cell or cell culture in which it was expressed. Contaminating components from its natural environment are materials that would generally interfere with the diagnostic or therapeutic uses of the polypeptide and may include enzymes, hormones, and other protein or non-protein solutes. In preferred embodiments, the antibody is purified (1) to a degree sufficient to yield at least 15 residues of internal or N-terminal amino acid sequence using a rotating beaker sequencer or (2) to homogeneity by SDS-PAGE under reduction or non-reduction conditions, using Coomassie blue staining or, preferably, silver staining. Isolated antibody includes antibodies in situ within recombinant cells, as at least one component of the polypeptide's natural environment is present.However, the isolated polypeptide is generally prepared by at least one purification step. The term "control sequences" refers to DNA sequences necessary for the operational expression of a linked coding sequence in a particular host organism. Control sequences suitable for prokaryotes, for example, include a promoter, optionally an operator sequence, and a ribosome binding site. RbCCnn / l 7Π7 / Β / Y Eukaryotic cells are known to use promoters, polyadenylation signals, and enhancers. A nucleic acid is "operationally bound" when it is in a functional relationship with another nucleic acid sequence. For example, the DNA for a presequence or secretory leader is operationally bound to the DNA of a polypeptide if it is expressed as a presequence protein involved in the secretion of the polypeptide; a promoter or enhancer is operationally bound to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operationally bound to a coding sequence if it is positioned to facilitate translation. In general, "operationally bound" means that the bound DNA sequences are contiguous and, in the case of a secretory leader, contiguous and in the reading frame. However, enhancers are not necessarily contiguous. Binding is achieved by attachment at convenient restriction sites.If these points do not exist, synthetic oligonucleotide adapters or linkers are used in accordance with conventional practice. “Percentage (%) amino acid sequence identity” with respect to the polypeptide sequences identified herein is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the sequence to which it is compared, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage of sequence identity, and without considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining the percentage of amino acid sequence identity can be achieved in various ways known to those skilled in the art, for example, using publicly available software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR).Experts in the technique can determine appropriate parameters for measuring alignment, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. For the purposes of this document, however, the amino acid sequence identity (%) values ​​are generated using the ALIGN-2 sequence comparison software. The ALIGN-2 sequence comparison software was created by Genentech, Inc., and the source code was filed with the user documentation with the U.S. Copyright Office, Washington, D.C. 20559, where it is registered under U.S. copyright registration number TXU510087. The ALIGN-2 program is publicly available from Genentech, Inc., South San Francisco, California. The ALIGN-2 program should be compiled for use on a UNIX operating system, preferably digital UNIX V4.0D.All sequence comparison parameters are set by the ALIGN-2 program and do not change. The amino acid sequences described herein are contiguous amino acid sequences, unless otherwise specified. RbCCnn / l 7Π7 / Β / Y "Structurally dissimilar" biological molecules, according to the present invention, refers to biological molecules that are not in the same class (protein, nucleic acid, lipid, carbohydrate, etc.) or, for example, when referring to proteins, having less than 60% amino acid identity, less than 50% amino acid identity, less than 40% amino acid identity, less than 30% amino acid identity, less than 20% amino acid identity, or less than 10% amino acid identity compared to each other. The expert in the technique can easily determine the "rigor" of the hybridization reactions, and this is generally an empirical calculation that depends on the probe length, wash temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes require lower temperatures. Hybridization generally depends on the ability of denatured DNA to re-hybridize when complementary strands are present in an environment below its melting temperature. The higher the desired degree of homology between the probe and the hybridizable sequence, the higher the relative temperature that can be used. As a result, it follows that higher relative temperatures tend to make the reaction conditions more rigorous, while lower temperatures make them less so.For further details and a rigorous explanation of hybridization reactions, refer to Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1995). "Strong conditions" or "highly stringent conditions", as defined herein, may be identified by: (1) use of low ionic strength and high temperature for washing, e.g., 0.015 M sodium chloride / 0.0015 M sodium citrate / 0.1% sodium dodecyl sulfate at 50C; (2) use during hybridization of a denaturing agent, such as formamide, e.g., 50% (v / v) formamide with 0.1% bovine serum albumin / 0.1% Ficoll.1% / 0.1% polyvinylpyrrolidone / 50 mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42 °C or (3) overnight hybridization in a solution using 50% formamide, 5x SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5x Denhardt's solution, ultrasonically homogenized salmon sperm DNA (50 pg / ml), 0.1% SDS and 10% dextran sulfate at 42 °C, with a 10-minute wash at 42 °C in 0.2x SSC (sodium chloride / sodium citrate) followed by a high-strength wash of 10 minutes of 0.1 x SSC with EDTA at 55 °C. “Moderately restrictive conditions” can be identified as described in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Coid Spring Harbor Press, 1989, and include the use of less stringent wash solution and hybridization conditions (e.g., temperature, ionic strength, and % SDS) than those described above. An example of moderately stringent conditions is overnight incubation at 37 °C in a solution comprising: 20% formamide, 5 x SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5 x Denhardt’s solution, 10% dextran sulfate, and 20 mg / ml of cut, denatured salmon sperm DNA, followed by washing the filters in 1 x SSC at approximately 37–50 °C. The expert in the technique will recognize how to adjust the temperature, ionic strength, etc. as needed to suit factors such as probe length and the like. Rbccnn / ι znz / R / v The “effector functions” of an antibody refer to the biological activities that can be attributed to the Fe region (a naturally occurring Fe region or an amino acid sequence variant Fe region) of an antibody and vary depending on the antibody isotype. Examples of antibody effector functions include: C1q binding and complement-dependent cytotoxicity; Fe receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor); and B cell activation. Antibody-dependent cell-mediated cytotoxicity (ADCC) refers to a form of cytotoxicity in which Ig secreted onto Fe receptors (FcRs) on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages) enables these cytotoxic effector cells to specifically bind to an antigen-bearing target cell and subsequently destroy the target cell with cytotoxins. Antibodies "load" the cytotoxic cells and are absolutely necessary for this destruction. The main cells mediating ADCC, NK cells, express only FcyRIII, whereas monocytes express FcyR1, FcyR2, and FcyR3. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991).To evaluate the ADCC activity of a molecule of interest, an in vitro ADCC assay can be performed, such as that described in U.S. Patent Nos. 5,500,362 or 5,821,337. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) lymphocytes. Alternatively or additionally, the ADCC activity of the molecule of interest can be evaluated in vivo, e.g., in an animal model, such as that described in Clynes et al. (Proc. Nati. Acad. Sci. U.S.A., 95:652-656 (1998)). "Fe receptor" or "FcR" describes a receptor that binds to the Fe region of an antibody. The preferred FcR is a naturally occurring human FcR. Furthermore, a preferred FcR is one that binds to an IgG antibody (a gamma receptor) and includes receptors of the FcyRI, FcyRII, and FcyRIII subclasses, including allelic variants and alternatively spliced ​​forms of these receptors. The FcyRII receptors include FcyRIIA (an "activating receptor") and FcyRIIB (an "inhibiting receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains. The activating receptor FcyRIIA contains an immunoreceptor tyrosine-based (ITAM) activation motif in its cytoplasmic domain. The inhibitory receptor FcyRIIB contains a tyrosine immunoreceptor-based inhibition motif (ITIM) in its cytoplasmic domain. Rev. Immunol. 15:203234 (1997)).The FcRs are discussed in Ravetch and Kinet (Annu. Rev. Immunol. 9:457-492 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995)). Other FcRs, including those that may be identified in the future, are encompassed by the term "FcR" herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994)). Rbccnn / ι ζηζ / E / γ Human effector cells are leukocytes that express one or more FcRs and perform effector functions. Preferably, the cells express at least FcγRIII and perform an ADCC effector function. Examples of human leukocytes that mediate ADCC include peripheral blood mononuclear cells (PBMCs), natural killer (NK) cells, monocytes, cytotoxic T lymphocytes, and neutrophils; PBMCs and NK cells are preferred. Effector cells can be isolated from a natural source, e.g., blood. Complement-dependent cytotoxicity (CDC) refers to the lysis of a target cell in the presence of complement. Activation of the classical complement pathway begins with the binding of the first component of the complement system (C1q) to antibodies (of the appropriate subclass) bound to its cognate antigen. To assess complement activation, a CDC assay can be performed, for example, as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). The term "therapeutically effective amount" refers to the quantity of an antibody or antibody fragment used to treat a disease or disorder in a subject. In the case of an allergic, inflammatory, or autoimmune disease (e.g., asthma, arthritis, etc.), the therapeutically effective amount of the antibody or antibody fragment (e.g., a multispecific or dual-specific antibody or antibody fragment for IL4 and IL5 or IL4 and IL13) may improve or treat the disease or prevent, reduce, improve, or treat symptoms associated with the disease. In the case of a proliferative disease (e.g.,In the case of a cancerous tumor, the therapeutically effective amount of the antibody or antibody fragment may reduce the number of cancer cells; reduce the size of the primary tumor; inhibit (i.e., slow down to some extent and preferably stop) the infiltration of cancer cells into peripheral organs; inhibit (i.e., slow down to some extent and preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate to some extent one or more of the symptoms associated with the disorder. To the extent that the antibody or antibody fragment can prevent the growth and / or destroy existing cancer cells, it may be cytostatic and / or cytotoxic.For cancer therapy, in vivo efficacy, for example, can be measured by assessing survival duration, time to disease progression (TTP), disease-free survival (DFS), progression-free survival (PFS), response rates (RR), duration of response, and / or quality of life. "Reduce or inhibit" indicates the ability to produce an overall decrease of preferably 20% or more, more preferably 50% or more, and most preferably 75%, 85%, 90%, 95%, or more. "Reduce" or "inhibit" may refer to the symptoms of the disorder being treated, the presence or size of metastases, the size of the primary tumor, or the size or number of blood vessels in angiogenic disorders. RbCCnn / l 7Π7 / Β / Y An "inflammatory disease," as used herein, refers to pathological states that produce inflammation, typically caused by neutrophil chemotaxis. An "autoimmune disease," as used herein, is a disease or disorder that arises from and is directed against an individual's own tissue or a co-secretion or manifestation thereof, or a condition resulting from it. Examples of inflammatory, autoimmune, or both diseases or disorders include, but are not limited to, asthma, such as bronchial asthma, bronchial asthma, and autoimmune asthma; arthritis (rheumatoid arthritis such as acute arthritis, chronic rheumatoid arthritis, gouty arthritis, acute gouty arthritis, chronic inflammatory arthritis, degenerative arthritis, infectious arthritis, Lyme arthritis, proliferative arthritis, psoriatic arthritis, vertebral arthritis, and juvenile-onset rheumatoid arthritis, osteoarthritis, chronic progressive arthritis, arthritis deformans, primary chronic polyarthritis, reactive arthritis, and ankylosing spondylitis); inflammatory hyperproliferative epithelial diseases; psoriasis, such as platelet psoriasis, guttate psoriasis, pustular psoriasis, and nail psoriasis; dermatitis, including contact dermatitis, chronic contact dermatitis, allergic dermatitis, allergic contact dermatitis, dermatitis herpetiformis, and atopic dermatitis;ax-linked hyper-IgM syndrome, urticaria, such as chronic allergic urticaria and chronic idiopathic urticaria, including chronic autoimmune urticaria, polymyositis / dermatomyositis, juvenile dermatomyositis, toxic epidermal necrolysis, scleroderma (including systemic scleroderma), sclerosis such as systemic sclerosis, multiple sclerosis (MS), such as spino-optic MS, primary progressive MS (PPMS) and relapsing-remitting MS (RRMS), progressive systemic sclerosis, atherosclerosis, arteriesclerosis, disseminated sclerosis and ataxic sclerosis, inflammatory bowel disease (IBD) (e.g. Crohn's disease, autoimmune-mediated gastrointestinal diseases, colitis such as ulcerative colitis, microscopic colitis, collagenous colitis, polyposis colitis, necrotizing enterocolitis and transmural colitis, as well as inflammatory bowel disease autoimmune), pyoderma gangrenosum, erythema nodosum, primary sclerosing cholangitis, episcleritis),respiratory distress syndrome, including acute respiratory distress syndrome (ARDS), meningitis, inflammation of all or part of the uvea, iritis, choroiditis, an autoimmune hematologic disorder, rheumatoid spondylitis, sudden hearing loss, IgE-mediated diseases such as anaphylaxis and atopic and allergic rhinitis, hyper-IgE syndrome, encephalitis such as Rasmussen's encephalitis and limbic and / or brainstem encephalitis, uveitis, such as anterior uveitis, acute anterior uveitis, granulomatous uveitis, non-granulomatous uveitis, phacoantigenic uveitis, posterior uveitis, or autoimmune uveitis, glomerulonephritis (GN) with and without nephrotic syndrome such as chronic or acute glomerulonephritis such as primary GN, immune-mediated GN, membranous GN (membranous nephropathy) idiopathic or idiopathic membranous nephropathy, membranous proliferative GN (MPGN), including type I and type II and rapidly progressing GN,allergic conditions, allergic reaction, eczema including atopic or allergic eczema, scleroderma, Whipple's disease, hypertrophic scarring, preeclampsia, abdominal adhesions, conditions involving T-cell infiltration and chronic inflammatory responses, chronic inflammatory lung disease, autoimmune myocarditis, leukocyte adhesion deficiency, systemic lupus erythematosus (SLE) or systemic lupus erythematosus erythematosus such as cutaneous SLE, cutaneous systemic lupus erythematosus, neonatal lupus syndrome (NLE), disseminated lupus erythematosus, lupus (including nephritis, cerebritis, pediatric, non-renal, extrarenal, RbCCnn / l 7P7 / B / Y discoid, alopecia) juvenile-onset diabetes mellitus (type I), including insulin-dependent diabetes mellitus (IDDM), adult-onset diabetes mellitus (type II diabetes), autoimmune diabetes, idiopathic diabetes insipidus, immune responses associated with acute or delayed hypersensitivity mediated by cytokines and T lymphocytes, tuberculosis, sarcoidosis, granulomatosis including lymphomatoid granulomatosis, Wegener's granulomatosis, agranulocytosis, vasculitis, including vasculitis (including large vessel vasculitis (including polymyalgia rheumatica and giant cell arteritis (Takayasu)), medium vessel vasculitis (including Kawasaki disease and polyarteritis nodosa), microscopic polyarteritis, CNS vasculitis, necrotizing, cutaneous or hypersensitive vasculitis, systemic necrotizing vasculitis and vasculitis associated with ANCA, such as Churg-Strauss vasculitis or Churg-Strauss syndrome (CSS), temporal arteritis, aplastic anemia,autoimmune aplastic anemia, Coombs-positive anemia, Diamond-Blackfan anemia, hemolytic anemia or immune hemolytic anemia including autoimmune hemolytic anemia (AIHA), pernicious anemia, Addison's disease, pure red cell aplasia or anemia (PRCA), factor VIII deficiency, hemophilia A, autoimmune neutropenia, pancytopenia, leukopenia, diseases involving leukocyte diapedesis, inflammatory disorders of the CNS, multiple organ injury syndrome such as those secondary to sepsis, trauma, or hemorrhage, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, antiphospholipid antibody syndrome, allergic neuritis, Behçet's disease, Castleman syndrome, Goodpasture syndrome, Raynaud's syndrome, Sjögren's syndrome, Stevens-Johnson syndrome, pemphigoid as bolus pemphigoid and epithelial pemphigoid, pemphigus (including pemphigus vulgar, pemphigus foliaceus,mucous membrane pemphigus (pemphigoid and pemphigus erythematosus), autoimmune polyendocrinopathies, Reiter's disease or syndrome, immune complex nephritis, antibody-mediated nephritis, neuromyelitis optica, polyneuropathies, chronic neuropathy such as IgM polyneuropathies or IgM-mediated neuropathy, thrombocytopenia (such as that which develops in patients with myocardial infarction, for example), including thrombotic thrombocytopenic purpura (TTP) and autoimmune or immune system-mediated thrombocytopenia such as idiopathic thrombocytopenic purpura (ITP) including chronic or acute ITP, autoimmune disease of the testes and ovaries, including autoimmune orchitis and oophthritis, primary hypothyroidism, hypoparathyroidism, autoimmune endocrine diseases including thyroiditis such as autoimmune thyroiditis, Hashimoto's disease, chronic thyroiditis (Hashimoto's thyroiditis), or subacute thyroiditis, autoimmune thyroid disease, idiopathic hypothyroidism, Graves' disease,polyglandular syndromes such as autoimmune polyglandular syndromes (or polyglandular endocrinopathy syndromes), paraneoplastic syndromes, including neurological paraneoplastic syndromes such as Lambert-Eaton myasthenic syndrome or Eaton-Lambert syndrome, muscle rigidity syndrome, encephalomyelitis such as allergic encephalomyelitis and experimental allergic encephalomyelitis (EAE), myasthenia gravis such as myasthenia gravis associated with thymoma, cerebellar degeneration, neuromyotonia, opsoclonus or opsoclonus-myoclonus (OMS) syndrome and sensory neuropathy, multifocal motor neuropathy, Sheehan's syndrome, autoimmune hepatitis, chronic hepatitis, lupoid hepatitis, giant cell hepatitis, chronic active hepatitis or autoimmune chronic active hepatitis, lymphoid interstitial pneumonitis, bronchiolitis obliterans (without transplantation) against NSIPs, Guillain-Barré syndrome, Berger's disease (nephropathy IgA), idiopathic IgA nephropathy, linear IgA dermatosis, primary biliary cirrhosis,pneumonocirrosis, autoimmune enteropathy syndrome, celiac disease, celiac sprue (gluten enteropathy), refractory sprue, idiopathic sprue, cryoglobulinemia, RbCCnn / l 7P7 / B / Y amylotrophic lateral sclerosis (ALS, Lou Gehrig's disease), coronary artery disease, autoimmune otic disease such as autoimmune inner ear disease (AIOD), autoimmune hearing loss, opsoclonus-myoclonus syndrome (OMS), polychondritis such as refractory or relapsing polychondritis, pulmonary alveolar proteinosis, amyloidosis, scleritis, a noncancerous lymphocytosis, a primary lymphocytosis including monoclonal B-cell lymphocytosis (e.g., benign monoclonal gammopathy and monoclonal gammopathy of undetermined significance, MGUS), peripheral neuropathy, paraneoplastic syndrome, channelopathies such as epilepsy, migraine, arrhythmia, muscle disorders, deafness, blindness, periodic paralysis and CNS channelopathies, autism, inflammatory myopathy, focal segmental glomerulosclerosis (FSGS), endocrine ophthalmopathy, uveoretinitis, chorioretinitis, autoimmune liver disorder, fibromyalgia,multiple endocrine failure, Schmidt's syndrome, adrenalitis, gastric atrophy, presenile dementia, demyelinating diseases such as autoimmune demyelinating diseases, diabetic nephropathy, Dressler's syndrome, alopecia areata, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia), autoimmune female and male infertility, mixed connective tissue disorder, Chagas disease, rheumatic fever, recurrent miscarriage, farmer's lung, erythema multiforme, postcardiotomy syndrome, Cushing's syndrome, poultry farmer's lung, allergic granulomatous angiitis, benign lymphocytic angiitis, Alport syndrome, alveolitis such as allergic alveolitis and fibrosing alveolitis, interstitial lung disease, transfusion reaction, leprosy, malaria, leishmaniasis, chianosomiasis, schistosomiasis, asariasis aspergillosis, Sampter syndrome, Caplan syndrome, dengue, endocarditis, endomyocardial fibrosis,diffuse interstitial pulmonary fibrosis, interstitial pulmonary fibrosis, idiopathic pulmonary fibrosis, cystic fibrosis, endophthalmitis, erythema elevatum diutinum, enthroblastosis fetalis, eosinophilic facilitis, Shulman syndrome, Felty syndrome, phlariasis, cyclitis such as chronic cyclitis, heterochronic cyclitis, iridocyclitis or Fuchs' cyclitis, Henoch-Schönlein purpura, human immunodeficiency virus (HIV) infection, echovirus infection, cardiomyopathy, Alzheimer's disease, parvovirus infection, rubella virus infection, post-vaccination syndromes, congenital rubella infection, Epstein-Barr virus infection, mumps, Evan syndrome, autoimmune gonadal failure, Sydenham's chorea, post-streptococcal nephritis, thromboangiitis media, thyrotoxicosis, tabes dorsalis, chorioretinitis, polymyalgia rheumatica, endocrine ophthalmopathy chronic hypersensitivity pneumonitis, keratoconjunctivitis sicca, epidermal keratoconjunctivitis,idiopathic nephritic syndrome, minimal change disease, benign familial ischemia-reperfusion injury, retinal autoimmunity, joint inflammation, bronchitis, chronic obstructive airway disease, silicosis, aphthous ulcers, aphthous stomatitis, arteriosclerotic disorders, aspermiogenesis, autoimmune hemolysis, Boeck's disease, cryoglobulinemia, Dupuytren's contracture, phacoanaphylactic endophthalmia, allergic enteritis, erythema nodosum leprosum, idiopathic facial paralysis, chronic fatigue syndrome, rheumatic fever, Hamman-Rich disease, sensorineural hearing loss, paroxysmal hemoglobinuria, hypogonadism, regional ileitis, leukopenia, infectious mononucleosis, transverse myelitis, primary idiopathic myxedema, nephrosis, sympathetic ophthalmia, granulomatous orchitis, pancreatitis, acute polyradiculitis, pyoderma gangrenosum, de Quervain's thyroiditis, acquired splenic atrophy, infertility due to antisperm antibodies,non-malignant thymoma, vitiligo, diseases associated with SCID and Epstein-Barr virus, acquired immune deficiency syndrome (AIDS), parasitic diseases such as leishmaniasis, toxic shock syndrome, RbCCnn / l 7P7 / B / Y food poisoning, conditions involving T-cell infiltration, leukocyte adhesion deficiency, immune responses associated with acute and delayed hypersensitivity mediated by cytokines and T lymphocytes, diseases involving leukocyte diapedesis, multiple organ injury syndrome, antigen-antibody complex-mediated diseases, antiglomerular basement membrane disease, allergic neuritis, autoimmune polyendocrinopathies, oophritis, primary myxedema, autoimmune atrophic gastritis, sympathetic ophthalmia, rheumatic diseases, mixed connective tissue disease, nephrotic syndrome, insulitis, polyendocrine failure, peripheral neuropathy, autoimmune polyglandular syndrome type I, adult-onset idiopathic hypoparathyroidism (IAHP), alopecia totalis, dilated cardiomyopathy, acquired epidermolysis bullosa (AEB), hemochromatosis, myocarditis, nephrotic syndrome, primary sclerosing cholangitis,purulent or non-purulent sinusitis, acute or chronic sinusitis, ethmoid, frontal, maxillary or sphenoid sinusitis, an autoimmune vesiculation disease, an eosinophil-related disorder such as eosinophilia, idiopathic hypereosinophilia syndrome, pulmonary infiltration eosinophilia, myalgia-eosinophilia syndrome, Löffler's syndrome, chronic eosinophilic pneumonia, tropical pulmonary eosinophilia, bronchopneumonic aspergillosis, aspergilloma or granulomas with eosinophils, anaphylaxis, seronegative spondyloarthritis, autoimmune polyendocrine disease, sclerosing cholangitis, sclerae, episcleras, chronic mucocutaneous candidiasis, Bruton's syndrome, transient hypogammaglobulinemia of childhood, Wiskott syndrome, Aldrich syndrome, ataxia telangiectasia, autoimmune disorders associated with collagen disease, rheumatism, neurological disease, ischemic reperfusion disorder, reduced blood pressure response, vascular dysfunction, angiectasia, tissue injury,Cardiovascular ischemia, hyperalgesia, cerebral ischemia and vascularization accompanying the disease, allergic hypersensitivity disorders, glomerulonephritis, reperfusion injury, reperfusion injury of the myocardium or other tissues, dermatoses with acute inflammatory components, acute purulent meningitis or other inflammatory disorders of the central nervous system, inflammatory disorders of the eyes or orbits, granulocyte transfusion-associated syndromes, cytokine-induced toxicity, severe acute inflammation, chronic intractable inflammation, pyelitis, pneumonocirrosis, diabetic retinopathy, diabetic disorders of large arteries, endartericular hyperplasia, peptic ulcer, valvulitis, and endometriosis. An "allergic disease" here refers to a disease or disorder in which an individual is hypersensitive and mounts an immune response to a substance that is not normally immunogenic. Allergic disease is generally characterized by the activation of mast cells via IgE, which produces an inflammatory response that can range from benign symptoms such as a runny nose to potentially life-threatening ones such as anaphylactic shock and death. Examples of allergic disease include, but are not limited to, asthma (e.g., allergic asthma), allergic rhinitis (e.g., hay fever), allergic dermatitis (e.g., eczema), contact dermatitis, food allergy, and urticaria. RbCCnn / l 7Π7 / Β / Y The above list is not exhaustive, and those skilled in the technique will understand that a disease or disorder may fall into several categories. For example, asthma is both an allergic and inflammatory disorder, and some physicians consider it an autoimmune disorder. The terms "cancer" and "cancerous" describe or refer to the physiological condition in mammals typically characterized by unregulated cell growth. This definition includes both benign and malignant cancers. The term "precancerous" refers to a condition or growth that typically precedes or ends in cancer. The term "proliferative disease," as used herein, refers to a disease or disorder associated with a certain degree of abnormal cell proliferation. In one modality, the proliferative cell disorder is cancer. In some modalities, the cancer is selected from the group consisting of breast cancer, colorectal cancer, non-small cell lung cancer, non-Hodgkin lymphoma (NHL), B-cell lymphoma, B-cell leukemia, multiple myeloma, kidney cancer, prostate cancer, liver cancer, head and neck cancer, melanoma, ovarian cancer, mesothelioma, and glioblastoma. The term "tumor," as used herein, refers to any growth and proliferation of neoplastic cells, whether malignant or benign, and to all precancerous and cancerous cells and tissues. "Non-metastatic" means a benign cancer that remains in its primary site and has not spread to the lymphatic system, blood vessels, or other tissues. Generally, a non-metastatic cancer is any stage 0, 1, or II cancer, and occasionally a stage III cancer. A "subject" is a vertebrate, preferably a mammal, more preferably a human being. Mammals include, but are not limited to, farm animals (such as cows), sport animals, pets (such as cats, dogs, and horses), primates, mice, and rats. II. Generation of specific double antibodies The heavy chain variable domain (Vh) contains considerably greater sequence diversity and contributes antigen recognition determinants more frequently than the light chain variable domain (Vl). Our previous results demonstrated, for the first time, that the light chain variable domain could be altered to produce a single antibody with dual specificity (see patent application no. 20080069820 and Bostrom et al., Science 232:1610-1614 (2009), which are incorporated herein by reference). We have discovered that dual-specific antibodies can be generated from antibodies that have Vl residues critical for antigen recognition by altering amino acid residues in the Vh, including the absence of mutations in the Vl. These novel and unexpected methods are described in greater detail below. RbCCnn / l 7Π7 / Β / Y We have discovered that specific residues in the Vl region can be identified which, when electrostatic or hydrophobic, suggest that the Vl region is crucial for antigen recognition. In these cases, alteration of the antibody's Vh region is used to generate a specific double-binding antibody that binds to both a first and a second epitope. Specifically, if any one, two, or three of the amino acid residues at positions 32, 50, or 91 (in Kabat numbering) of an antibody are electrostatic (e.g., tyrosine) or hydrophobic (e.g., tryptophan), then the nucleic acid sequence encoding the Vh region of that antibody is altered in one or more Vh codons that encode one or more solvent-accessible amino acid residues. In various forms, the residues in the Vh can be altered to include any one or more of the amino acids 33, 34, 50-58, or 95-97. Optionally, amino acid residues 93-96 of the light chain can be altered in addition to the heavy chain residues. Solvent accessibility or importance for antigen recognition can be determined for the heavy chain using standard techniques known in the field, including, but not limited to, structural matching and alanine scanning mutagenesis. The altered Vl and Vh are then expressed (e.g., as a gene library), and a specific double antibody or antigen-binding fragment thereof is selected, containing an altered Vl and Vh that binds specifically to a first and second epitope. The Vl of the initial antibody may or may not be altered in addition to the Vh. The amino acid residues in the frame region of the initial antibody may or may not be altered in addition to the Vh. A specific double antibody or antigen-binding fragment identified by the above selection methods can be further modified, for example, by affinity maturation or other methods known in the art, to increase affinity for one or both target antigens. The affinity maturation selection process may include the application of a massively parallel sequencing approach (e.g., deep sequencing, ultra-deep sequencing, or next-generation sequencing) to identify one or more residues (e.g., solvent-exposed or solvent-unexposed residues) that contribute to the binding of one or both target antigens (e.g., contribute to increasing affinity for the target antigen). See, for example, Fowler et al. Nat. Methods. 7(9): 741-746, 2010. The specific double antibody can also be modified to increase stability or half-life or to decrease immunogenicity.The expert in the technique is aware of these modifications. III. Therapeutic uses The specific double antibodies or antigen-binding fragments described herein that bind to both IL4 and IL5 (e.g., mature affinity variants Β1, E7 and E7) or IL4 and IL13 (e.g., F1 and F2) can be used to treat, suppress or prevent diseases, such as allergic, inflammatory and autoimmune diseases (e.g., asthma), IL4-mediated diseases, IL5-mediated diseases, IL13-mediated diseases, IL4 / IL5-mediated diseases, IL4 / IL13-mediated diseases and / or proliferative disorders (e.g., cancer). RbCCnn / l 7Π7 / Β / Y Examples of inflammatory and autoimmune diseases or disorders that can be treated with specific double-stranded antibodies or antigen-binding fragments of these were described above. In some modalities, the disease or disorder includes, but is not limited to, asthma such as bronchial asthma, bronchial asthma, and autoimmune asthma. Asthma is described as a chronic lung disease involving airway inflammation, hyperresponsiveness, or obstruction. Physiologically, airway hyperresponsiveness is documented by decreased bronchial airflow following bronchoprovocation with methacholine or histamine. Other triggers that cause airway obstruction include cold air, exercise, viral upper respiratory infection, smoking, and respiratory allergens. Bronchial provocation with allergens involves a rapid, early-phase, immunoglobulin E (IgE)-mediated decrease in bronchial airflow, followed in many patients by a late-phase, IgE-mediated reaction with decreased bronchial airflow for 4–8 hours.The early response is triggered by the acute release of inflammatory substances, such as histamine, PGD2, leukotrienes, tryptase, and platelet-activating factor (PAF), while the late response is triggered by de novo synthesized proinflammatory cytokines (e.g., TNFα, IL4, IL13) and chemokines (e.g., MCP-1 and MIP-1α) (Busse et al. In: Allergy: Principles and Practice, Ed. Middleston, 1173 (1998)). In chronic asthmatic patients, persistent pulmonary symptoms are mediated by the superior Th2 cell response. Th2 cytokines are believed to have a vital impact on the disease (Larche et al., J. Allergy Clin. Immunol., 111: 450 (2003)), in particular IL13 and IL4 produced by Th2 cells with the NK phenotype (NKT) in the airways as indicated in a rodent asthma model (Akbari et al., Natura Med., 9: 582 (2003)).The macroscopic pathology of asthmatic airways presents with pulmonary hyperinflammation, smooth muscle hypertrophy, reticular lamina thickening, mucosal edema, epithelial cell detachment, ciliary cell dysfunction, and mucus gland hypersecretion. Microscopically, asthma is characterized by an increased number of eosinophils, neutrophils, lymphocytes, and plasma cells in bronchial tissues, bronchial secretions, and mucus. Initially, leukocytes are recruited from the bloodstream to the airways by activated CD4+ T lymphocytes. Activated T lymphocytes also direct the release of inflammatory mediators from eosinophils, mast cells, and lymphocytes. Additionally, Th2 cells produce IL-4, IL-5, IL-9, and IL-13. IL-4, along with IL-13, signals the exchange of IgM antibodies for IgE. The cross-linking of membrane-bound IgE molecules by allergens triggers mast cell degranulation, which releases histamine, leukotrienes, and other mediators that perpetuate airway inflammation. IL-5 activates eosinophil recruitment and activation. Activated mast cells and eosinophils also generate their own cytokines, which further contribute to the inflammation. These repeated cycles of inflammation in the lungs, with lung tissue injury followed by repair, can lead to long-term structural changes ("remodeling") of the airways. RbCCnn / l 7Π7 / Β / Y Moderate asthma is currently treated with anti-inflammatory corticosteroids or a mast cell inhibitor such as cromolyn sodium or nedocromil plus an inhaled beta-2 agonist as needed (3–4 times daily) to relieve symptoms of an exacerbation or asthma induced by allergens or exercise. Cromolyn sodium and nedocromil block bronchospasm and inflammation, but are usually effective only for asthma associated with allergens or exercise and typically only for young asthmatics. Inhaled corticosteroids improve inflammation, airway hyperreactivity, and obstruction, and reduce the number of acute exacerbations. However, at least nine months are required before effects are visible and up to a year for noticeable improvement. The most common side effects are hoarseness and oral fungal infection, i.e., candidiasis. More serious side effects have been reported, e.g.For example, partial adrenal suppression, growth inhibition, and decreased bone formation, but only with the use of higher doses. Beclomethasone, triamcinolone, and flunisolide likely have similar potency, while budesonide and fluticasone are more potent and reportedly have fewer systemic side effects. Even patients with mild disease exhibit airway inflammation, including infiltration of the mucosa and epithelium with activated T cells, mast cells, and eosinophils. T cells and mast cells release cytokines that promote eosinophil growth and maturation and IgE antibody production. These antibodies, in turn, increase microvascular permeability, disrupt the epithelium, and mimic neural reflexes and mucus-secreting glands. The result is airway hyperreactivity, bronchoconstriction, and hypersecretion, manifested by sneezing, coughing, and dyspnea. Traditionally, asthma has been treated with oral and inhaled bronchodilators. These agents help with asthma symptoms but do nothing for the underlying inflammation. Recognition over the past 10 years of the importance of inflammation in the etiology of asthma has led to increased use of corticosteroids, but many patients continue to suffer from uncontrolled asthma. Specific double antibodies or antigen-binding fragments of these with specificity for IL4, IL5 and / or IL13 would target multiple pathogenic pathways and can be used as a therapeutic agent, alone or in combination with additional therapies (e.g., those known in the technique or described above), for the treatment of asthma. In additional modalities, specific double-stranded antibodies or antigen-binding fragments of these antibodies can be used to treat cancer. The term cancer encompasses a collection of proliferative disorders that include, but are not limited to, precancerous growths, benign tumors, and malignant tumors. Benign tumors remain localized at their site of origin and do not have the ability to infiltrate, invade, or metastasize to distant sites. Malignant tumors invade and damage other tissues surrounding them. They can also acquire the ability to break away from their site of origin and spread to other parts of the body (metastasize), usually through the bloodstream or via the lymphatic system where lymph nodes are located. Primary tumors are classified according to the type of tissue from which they arise; metastatic tumors are classified according to the type of tissue from which the cells are derived. RbCCnn / l 7P7 / B / Y cancerous. Over time, the cells of a malignant tumor become more abnormal and look less like normal cells. This change in the appearance of cancer cells is called tumor grade, and cancer cells are described as well-differentiated, moderately differentiated, poorly differentiated, or undifferentiated. Well-differentiated cells look fairly normal and resemble the normal cells from which they originated. Undifferentiated cells are cells that have become so abnormal that it is no longer possible to determine their origin. IV. Doses and formulations The antibody or antibody fragment compositions shall be formulated, dosed, and administered in a manner consistent with good medical practice. Factors to be considered in this context include the specific disorder being treated, the specific mammal being treated, the individual patient's clinical condition, the cause of the disorder, the site of administration of the agent, the method of administration, the schedule of administration, and other factors known to medical professionals. The "therapeutically effective amount" of the antibody or antibody fragment to be administered shall be governed by these considerations and is the minimum amount necessary to prevent, ameliorate, or treat a disease or an allergic, inflammatory, autoimmune, or proliferative disorder, or symptoms thereof.The dosage and timing of administration of the antibody or antibody fragment of the invention will depend on various clinical factors, including the subject's general health and the severity of symptoms, for example, of the allergic disorder. The invention includes the use of antibodies or antibody fragments to treat, prevent, or reduce allergic disorders or their symptoms, or the risk of developing allergic disorders in a subject. The antibody or antibody fragment can be administered at any time, for example, after the diagnosis or detection of an allergic disorder or a condition associated with an allergic disorder, or for the prevention of an allergic disorder in subjects who have not yet been diagnosed with an allergic disorder but are at risk of developing it (e.g.,, subjects who suffer from or have received treatment for a compromised immune system), after determining a risk of developing an allergic disorder. The specific double antibodies or antigen-binding fragments thereof of the present invention can be formulated and administered in various forms, e.g., known routes for specific indications, including, but not limited to, inhalation, topical, oral, subcutaneous, bronchial injection, intravenous, intracerebral, intranasal, transdermal, intraperitoneal, intramuscular, intrapulmonary, vaginal, rectal, intra-arterial, intracerebrospinal, intra-articular, intrasynovial, intralesional, parenteral, intraventricular in the brain, or infraocular. For example, the antibodies or antibody fragments may be in the form of a pill, tablet, capsule, liquid, or sustained-release tablet for oral administration, or a liquid for intravenous or subcutaneous administration, a polymer or other sustained-release vehicle for local administration, an ointment, cream, gel, liquid, or patch for topical administration. Rbccnn / ι ζηζ / E / γ Local administration may be particularly desirable if extensive side effects or toxicity are associated with IL-4, IL-5, or IL-13 antagonism. An ex vivo strategy can also be used for therapeutic applications. Ex vivo strategies involve transfecting or transducing cells obtained from the subject with a polynucleotide encoding a specific double-stranded antibody or antigen-binding fragment. The transfected or transduced cells are then returned to the subject. The cells can be any of a broad spectrum of types, including, but not limited to, hematopoietic cells (e.g., bone marrow cells, macrophages, monocytes, dendritic cells, T cells, or B cells), fibroblasts, epithelial cells, endothelial cells, keratinocytes, or muscle cells. For example, continuous systemic infusion or periodic injection of specific double antibodies or antigen-binding fragments thereof can be used to treat or prevent the disorder. Treatment can continue for a period ranging from 1 day to the rest of the subject's life, more preferably 1 to 100 days and even more preferably 1 to 20 days, most preferably until the symptoms of the allergic, inflammatory, autoimmune, or proliferative disease or disorder are reduced or eliminated. Dosages vary depending on the compound and the severity of the condition. Specific double antibodies or antigen-binding fragments thereof can be administered continuously by infusion, using an implantable constant-flow or programmable pump, or by periodic injections. Sustained-release systems can also be used.Semipermeable implantable membrane devices are also useful as delivery methods for specific double antibodies or antigen-binding fragments thereof under certain circumstances. In another modality, specific double antibodies or antigen-binding fragments thereof are administered locally, e.g., by inhalation, and can be repeated periodically. Pulmonary administration can also be employed, e.g., using an inhaler or nebulizer and the formulation with an aerosol-forming agent. The antibody can also be delivered to a patient's lungs in the form of a dry powder composition (refer, e.g., to U.S. Patent No. 6,514,496). The dosage of specific double antibodies or antigen-binding fragments thereof will depend on other clinical factors such as the subject's weight and condition, as well as the route of administration. For treating subjects, doses of specific double antibodies or antigen-binding fragments thereof can be administered at approximately 0.1 mg / kg to 500 mg / kg of body weight. A more preferred range is 1 mg / kg to 50 mg / kg of body weight, with the most preferred range being between 1 mg / kg and 25 mg / kg of body weight. Depending on the half-lives of the antibodies or antibody fragments in the particular subject, the antibodies or antibody fragments can be administered several times a day to once a week. The methods of the present invention provide for single as well as multiple administrations, delivered simultaneously or over a prolonged period. Preferably, specific double antibodies or antigen-binding fragments thereof are administered parenterally or intravenously by continuous infusion or locally via inhaler. The dose and regimen of RbCCnn / l 7P7 / B / Y dosages depend on the severity of the disease and the subject's general health. For parenteral administration, specific double antibodies or antigen-binding fragments thereof are formulated as a unit-dose injectable form (solution, suspension, emulsion) in combination with a pharmaceutically acceptable parenteral vehicle. These vehicles are inherently non-toxic and non-therapeutic. Examples of such vehicles include water, saline solution, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous vehicles, such as non-volatile oils and ethyl oleate, may also be used. Liposomes may also be used as carriers. The vehicle may include small amounts of additives such as substances that enhance isotonicity and chemical stability, e.g., buffers and preservatives.Specific double antibodies or antigen-binding fragments thereof are typically formulated in these vehicles at concentrations of approximately 1 mg / ml to 10 mg / ml. For administration by inhalation, specific double antibodies or antigen-binding fragments thereof are formulated by any suitable method to form the aerosol form of the present invention. A composition comprising specific double antibodies or antigen-binding fragments thereof is vaporized or nebulized with or without a pharmaceutically acceptable excipient to produce a vapor that can be condensed and inhaled directly into the lungs using, for example, an inhaler. Pharmaceutically acceptable excipients may be volatile, and classes of such excipients are known in the art and include, but are not limited to, gaseous supercritical fluid, liquid solvents, and solid solvents.Examples of carriers within the classes include, but are not limited to, water, sterile saline solution, physiological buffer solutions such as phosphate-buffered saline, terpenes, alcohols, propylene glycol, glycerol and other similar alcohols, dry ice, dimethylformamide, dimethylacetamide, supercritical carbon dioxide and mixtures thereof. The required dosage depends on the chosen route of administration, the nature of the formulation, the nature of the subject's illness, the subject's size, weight, surface area, age, and sex, other drugs being administered, and the treating physician's judgment. Wide variations in the required dosage should be expected depending on the variety of polypeptides and fragments available and the different efficacies of the various routes of administration. For example, oral administration would be expected to require higher dosages than administration by intravenous injection. Variations in these dosage levels can be adjusted using standard empirical routines for optimization, as is known in the art. Administrations can be single or multiple (e.g., 2, 3, 6, 8, 10, 20, 50, 100, 150, or more). Encapsulation of the polypeptide in a suitable delivery vehicle (e.g.,, polymeric microparticles or implantable devices) can increase the effectiveness of delivery, particularly for oral delivery. In one modality for the treatment of asthma, it is optional, not required, that the specific double antibody or antigen-binding fragment be formulated or administered in conjunction (simultaneously or sequentially) with one or more agents currently used to prevent or treat asthma or the risk of developing asthma. The antibody or antigen-binding fragment can be formulated, for example, with an IgE antagonist or other drugs. RbCCnn / l 7P7 / B / Y bronchodilators (e.g., a beta2-adrenoceptor agonist, xanthines, cholinesterase antagonists), anti-inflammatory agents (e.g., cromolyn sodium (DSCG), nedocromil sodium, antihistamines such as ketotifen, corticosteroids such as prednisolone), theophylline, salbutamol, beclomethasone dipropionate, or another asthma therapeutic agent known in the art. The effective amount of these additional agents depends on the amount of specific double-stranded antibody or antigen-binding fragment of it present in the formulation, the type of disorder or treatment, and other factors mentioned above. Therapeutic formulations are prepared using standard methods known in the art by mixing the active ingredient with the desired degree of purity with optional physiologically acceptable carriers, excipients, or stabilizers (Remington's Pharmaceutical Sciences (20th edition), ed. A. Gennaro, 2000, Lippincott, Williams & Wilkins, Philadelphia, PA).Acceptable carriers include saline solution or buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight polypeptides (less than approximately 10 residues); proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN™, PLURONICS™, or PEG. Optionally, but preferably, the formulation contains a pharmaceutically acceptable salt, preferably sodium chloride, and preferably at approximately physiological concentrations. Optionally, the formulations of the invention may contain a pharmaceutically acceptable preservative. In some embodiments, the preservative concentration ranges from 0.1 to 2.0%, typically v / v. Suitable preservatives include those known in pharmaceutical techniques. Benzyl alcohol, phenol, m-cresol, methylparaben, and propylparaben are examples of preservatives. Optionally, the formulations of the invention may include a pharmaceutically acceptable surfactant at a concentration of 0.005 to 0.02%. The formulation herein may also contain more than one active compound as required for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Such molecules are present in appropriate combinations in amounts that are effective for the desired purpose. The active ingredients can also be encapsulated in prepared microcapsules, for example, using coacervation techniques or interfacial polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively; in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules); or in macroemulsions. RbCCnn / l 7Π7 / Β / Υ techniques are described in Remington's Pharmaceutical Sciences, referred to above. Sustained-release preparations can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which are in the form of molded articles, e.g., films or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactides (U.S. Patent No. 3,773,919), L-glutamic acid and ethyl-L-glutamate copolymers, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid.While polymers such as ethylene vinyl acetate and glycolic-lactic acid allow for the release of molecules for more than 100 days, certain hydrogels release proteins for shorter periods. When encapsulated antibodies remain in the body for an extended time, they can denature or accumulate as a result of exposure to humidity at 37°C, leading to a loss of biological activity and potential changes in immunogenicity. Rational stabilization strategies can be devised depending on the mechanism involved. For example, if the accumulation mechanism is found to be the formation of intermolecular SS bonds via thio-disulfide exchange, stabilization can be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions. In one example, the specific double antibody or antigen-binding fragment thereof is administered locally, e.g., by direct injection when the disorder permits, and the injections can be repeated periodically, or by inhalation. The specific double antibody or antigen-binding fragment thereof can also be delivered systemically to the subject or directly to the affected area. The invention also provides a composition comprising the specific double antibody or antigen-binding fragment thereof and a pharmaceutically acceptable carrier or diluent. This composition for therapeutic use is sterile and lyophilizable. The use of a specific double antibody or antigen-binding fragment thereof of the present invention is also contemplated in the manufacture of a medicament for the treatment of an indication described herein. The composition may also comprise a second therapeutic agent such as an anti-asthmatic agent, an anti-inflammatory agent, or an antiproliferative agent (e.g., a chemotherapeutic agent, a cytotoxic agent, or an antiangiogenic agent). RbCOnn / l 7Π7 / Β / Y V. Manufacturing kits and items Another embodiment of the invention is a manufactured article containing materials useful for treating diseases or disorders (e.g., allergic diseases or disorders, or asthma). Yet another embodiment of the invention is a manufactured article containing materials useful for treating inflammatory, autoimmune, and proliferative diseases or disorders. The manufactured article comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, flasks, syringes, etc. The containers may be formed from various materials, such as glass or plastic. The container holds a composition that is effective in treating the condition and may have a sterile access port (e.g., the container may be a bag or intravenous solution bottle with a stopper that can be pierced by a hypodermic injection needle).At least one active agent in the composition is a dual-specific antibody or antigen-binding fragment of the invention. The label or package insert indicates that the composition is used in the treatment of the particular condition. The label or package insert shall also include instructions for administering the antibody composition to the patient. Articles of manufacture and kits comprising combination therapies described herein are also contemplated. The package insert refers to instructions typically included in commercial packaging of therapeutic products, containing information about the indications, use, dosage, administration, contraindications, and / or warnings related to the use of those products. In one instance, the package insert indicates that the composition is used to treat asthma. Additionally, the manufactured article may further comprise a second container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and dextrose solution. It may also include other commercially and user-desirable materials, including additional buffers, diluents, filters, needles, and syringes. Kits are also provided for various purposes, e.g., for the purification or immunoprecipitation of IL4, IL5, or IL13 from cells. For the isolation and purification of IL4, IL5, or IL13, the kit may include an IL4 / IL5 or IL4 / IL13 antibody coupled to microspheres (e.g., Sepharose microspheres). Kits containing the antibodies for the in vitro detection and quantification of IL4, IL5, or IL13 may be provided, e.g., in an ELISA or Western blot assay. As a manufactured item, the kit comprises a container and a label or package insert on or associated with the container. The container holds a composition comprising at least one specific bispecific or multispecific antibody or antibody fragment of the invention. Additional containers may be included that hold, e.g., diluents and buffers or control antibodies.The label or package leaflet may provide a description of the composition, as well as instructions for in vitro or intended diagnostic use. The following examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way. In fact, various modifications of the invention, in addition to those shown and described herein, will be evident to those skilled in the art from the foregoing description and will be found Rbccnn / ι ζηζ / E / γ within the scope of the attached claims. EXAMPLES The commercially available reagents mentioned in the examples were used according to the manufacturer's instructions, unless otherwise indicated. The origin of the cells identified in the following examples and throughout this specification by their ATCC registration numbers is the American Type Culture Collection, Manassas, VA. Unless otherwise indicated, the present invention utilizes standard recombinant DNA technology procedures, as described above and in the following bibliography: Sambrook et al., mentioned above; Ausubel et al., Current Protocols in Molecular Biology (Green Publishing Associates and Wiley Interscience, NY, 1989); Innis et al., PCR Protocols: A Guide to Methods and Applications (Academic Press, Inc.: NY, 1990); Harlow et al., Antibodies: A Laboratory Manual (Coid Spring Harbor Press: Coid Spring Harbor, 1988); Gait, Oliqonucleotide Synthesis (IRL Press: Oxford, 1984); Freshney, Animal Cell Culture, 1987; Coligan etál., Current Protocols in Immunology, 1991. Ejemplo 1. Diseño y construcción de genoteca Through mutation in the light chain (LC) complementarity-determining regions (CDRs), a monospecific antibody can acquire a second binding specificity for a novel antigen while maintaining its primary antigen specificity. Furthermore, dual binding can occur with affinity maturation down to a low nanomolar Kd. This pathway for dual specificity modification is well-suited for antibodies that primarily utilize heavy chain (HC) CDRs for binding to their primary antigens. Here, we describe an anti-interleukin-4 antibody derived from a mouse hybrid, which is highly dependent on LC CDRs for binding energy, and the evolution of this antibody to dual specificity through mutation only in the HC CDRs or with an additional mutation in the LC CDRs.A specific dual variant of this type with respect to interleukin 4 (IL4) and interleukin 5 (IL5) matures to high dual affinity with a low nanomolar Kd. The results further highlight the antibody's ability to evolve dual specificity and demonstrate a generally applicable modification circuit for evolving dual specificity in any antibody using a mutation-scanning strategy to identify regions of an antigen-binding site that can tolerate mutation and allow the evolution of secondary binding specificity. We aimed to recruit a second antigen specificity to a humanized 19C11 antibody (hu19C11, humanized at the germline base structure VH1 and kappa I), which binds to IL4 and blocks its binding to the IL4α receptor. First, its Fab was cloned into a phagemid construct (pV0115) to co-express the LC and, bicistronally, the variable domain and constant domain 1 (CH1) of HC fused at its C-terminus to the minor coating protein M13p3 as described (Lee et al., J. Mol. Biol. 340:1073-93, 2004). A portion of the IgG hinge region with the amino acid sequence KTHTC was included between CH1 and M13p3 to allow the expression of RbCCnn / l 7Π7 / Β / Y Bivalent Fab (Lee et al., J. Mol. Biol. 340:1073-93, 2004) was used to increase the binding efficiency to antigens immobilized on a solid surface support. First, it was verified that the Fab-expressing phage bound well to an antibody against the expression tag (gD) fused at the C-terminus to LC and IL4 with high affinity (phage ECs = 1 nM), which is similar to the measured affinity of the antibody as IgG. To determine the recruitment modification strategy for a secondary binding specificity with respect to hu19C11, the importance of its three CDR LCs for IL4 binding was first examined by mutating two or three residues of each CDR LC to alanine. These CDR LC positions were selected due to their previous identification as key positions for combinatorial mutagenesis in the LC library approach to selecting specific double clones from antibodies that heavily utilize the CDR HCs for their major antigen binding (Bostrom et al. PLoS One. 6:e17887, 2011). Wild-type hu19C11-expressing phages, alanine mutants L1 (I30A / N31A / D32A), L2 (Y50A / H53A / R54A), or L3 (D91A / Y92A), were immobilized in anti-gD antibody (Figure 1A) or IL4 (Figure 1B) and analyzed using ELISA to determine binding. CDR LCs were found to be energetically relevant.Phages expressing Fab with alanine mutations in any of the three CDR LCs did not exhibit detectable IL4 binding, but they did bind to anti-gD antibody, indicating that these mutants were indeed present in the phages, albeit at a lower level than wild-type Fab, but their IL4 binding was significantly altered (Figure 1A and 1B). Therefore, the approach of generating randomly separated CDR LC libraries is unlikely to produce double-specific antibodies, as the number of amino acid residues available for secondary antigen-binding evolution is considerably limited. Key individual CDR LC residues, as well as surface-accessible CDR HCs, were subsequently mutated as previously described (Sidhu et al. J Mol Biol. 338:299-310, 2004; Lee et al. J Mol Biol.340:1073-1093, 2004) and its impact on IL4 binding was assessed by determining the relative binding affinity of these alanine mutants compared to wild-type hu 19C11. The results confirmed the importance of key CDR LC residues for IL4 binding. However, all analyzed positions of CDR H2 and half of CDR H1 and H3 appear to be tolerant to the mutation (Figure 1C). By mapping the critically important residues for IL4 binding (LC residues 30, 31, 32, 50, 53, 54, 91, 92; HC residues 31, 32, 98, 99 (in Kabat numbering)) to the Fab structure of trastuzumab (PDB: 1FDV) as a model, a centralized area in CDR H2 was identified that tolerated the mutation and could be suitable for evolving a second specificity (Figure 2A). A set of residues in CDR H1 (33, 34), H2 (50-58), H3 (95-97) and L3 (93-96) that tolerated alanine mutation was then selected to centralize random separation in CDR H2 for the mutation scheme and a set of synthetic oligonucleotides was used to randomly separate these selected residues in each of the four CDRs according to the mutagenesis method of Kunkel et al. (Kunkel et al., Methods Enzymol. 154:367-82, 1987).The randomization scheme was guided by the diversity in natural antibodies in terms of amino acid composition and the extent of variability in CDR H2 and CDR L3 (Figure 2B). The template for mutagenesis included a stop codon only in CDR H2. Rbccnn / ι znz / R / Y ensured the mutation in this CDR in Fab-expressing libraries. Ten phage expression libraries were generated, each with sizes ranging from 108-109. As assessed by hu19C11 phage titers and the ten HC libraries (2144-1 to 2144-10), the binding to immobilized IL4 (PeproTech) captured by an unblocked anti-IL4 antibody, each library, as a whole, showed a certain low level of IL4 binding that was generally greatly reduced compared to the hu19C11 template antibody, indicating different levels of alteration of average IL4 binding (Figure 3). IL5 and IL13 were selected as secondary antigens. Although these two interleukins, like IL4, belong to the quadrupole cytokine family, they are considerably divergent at the amino acid sequence level and structural organization (LaPorte et al. Cell. 132:259-272, 2008; Patino et al. Structure. 19:1864-1875, 2011; Finkelman et al. J Immunol. 184:1663-1674, 2010). The sequence identity between IL4 and IL13 is 12%, while that between IL4 and IL5 is 11%. At the structural level, IL5 forms a unique cross-linked homodimer with one α helix from one chain forming a group of 4 helices with three α helices from the other chain (Milburn et al. Nature. 363:172-176, 1993; Patino et al. Structure. 19:1864-1875, 2011); IL4 and IL13 are both monomers.However, the three cytokines are related in their biological function and specific double antibodies that bind to and block two of these cytokines may have potential use as treatments for allergic diseases such as asthma 9 (Finkelman et al. J Immunol. 184:1663-1674, 2010; Haldar et al. NEngIJMed. 360:973-984, 2009). Several rounds of adsorption and enrichment were performed for IL5- or IL13-binding clones with IL4-binding retention from phage expression libraries constructed as previously described (Bostrom et al. Science. 323:1610-1614, 2009). Screening approximately 100 clones of each gene yielded 3 to 8 clones exhibiting dual IL5 / IL4 or IL13 / IL4 binding, respectively. Sequencing identified two unique IL4 / IL5-binding clones (B1, E7) and two unique IL4 / IL13-binding clones (F1, F2) (Figure 2B). IL5-binding-only clones (e.g., clone 5A) were also isolated. With the exception of clone B1, all clones included mutations only in the HC CDRs compared to their original monospecific template (Figures 4A-4C). This demonstrated that the CDR HC mutation of a monospecific antibody can confer dual specificity.Phage binding assays were used to calculate clone affinity as the interleukin concentration required to inhibit 50% of Fab expression on phages from binding to immobilized interleukins (IC50). Binding to secondary antigens was found to be weak with IC50 in the micromolar range, while IL4 binding remained in the low nanomolar range. Example 2. Evaluation of the performance of gene libraries To verify double-binding specificity, clones B1, E7, F2, and 5A were expressed as IgG, and the binding of these resulting IgGs to the expressed antigens was confirmed, but not to various other proteins (Figure 5). Binding specificity was also analyzed by demonstrating minimal IgG binding to cells of the human epithelial kidney cell line 293 Rbccnn / ι znz / R / v by flow cytometry, without expression of IL4, IL5 and IL13, as well as to baculovirus (BV) particles generated from insect cell lines by ELISA (Hotzel et al. MAbs. 4:753-760, 2012). Additionally, the specific double IL4 / IL5 antibodies B1 and E7 together with the monospecific IL5-binding antibody showed to block the binding of IL5 to the IL5 receptor or of IL5 itself, suggesting that the binding epitopes on IL5 overlap with those of the IL5 receptor (Figure 6). Through surface plasmon resonance (SPR) measurements, the specific double IL4 / IL5 clone had low affinity with respect to IL5 (Kd = 905 nM), but maintained high affinity in binding to IL4 (Kd = 3.4 nM) of its parent antibody hu19C11, as measured by surface plasmon resonance (SPR) (Figures 7A and 7B). Example 3. Affinity maturation of the double specific antigen-binding fragment To improve the dual affinity of E7, CDR E7s were randomly separated by site-directed mutagenesis and the variants were presented to phages for binding selection. Three libraries were generated by targeting CDR H2 and CDR L3 residues (H2 / L3 library), CDR H1, H2 and H3 residues (H1 / H2 / H3 library), or CDR H2 residues and selected points in the frame 3 (FR3) region of HC (H2 / FR3 library) for random separation as described (Lee et al. J Mol Biol. 340:1073-1093, 2004; Bostrom et al. Methods Mol Biol. 525:1-24, 2009; Lee et al. Blood. 108:3103-3111, 2006) (Figure 7A). Since clone E7 maintained a high IL4 binding affinity, the gene library selection was focused on improving IL5 binding.From the H1 / H2 / H3 library, many clones with improved affinity for IL5 but significantly reduced affinity for IL4 were discovered, while clones from the H2 / L3 and H2 / FR3 libraries showed improved IL5 binding without loss of IL4 binding affinity (Figure 4A). Selected clones were purified as IgG and compared to E7. Many H2 / L3 library variants exhibited increased double binding as IgG without increased binding to a set of non-target proteins (Figure 8A). Low off-target binding was also confirmed using BV and FACS binding from 293 cells, as previously described, and the improved clones continued to block IL5 binding to its receptor (Figure 8B). The monovalent binding affinities of the two enhanced variants, 1C36 and 1C60 (Figures 7C and 7D), were determined through SPR measurements as Fabs binding to immobilized IL4 or IL5.Both variants maintained a high affinity with respect to IL4 (Kd = 3-4 nM) and better affinity with respect to IL5 of 37 times and 20 times respectively (Kd = 24.1 nM for 1C36 and 44.4nM for 1C60) (Figure 7B). In summary, it was shown that mutation in the CDR HC of a monospecific antibody can recruit a second binding specificity, and one of the dual-specific antibodies was enhanced to a low nM affinity for both antigens. Previously, trastuzumab Fab was shown to evolve dual specificity through mutation in the CDR LC; since then, other dual-specific antibodies have been generated using the same LC approach. Together with the findings of the current study, the capacity for evolution of antibody binding specificity is highlighted. It was discovered that with limited mutation in CDRs not only on the light chain side but also on the heavy chain side, an antibody can add another binding specificity. In nature, antibodies are constantly being remodeled through gene transposition and somatic mutation. It was observed that antibodies can RbCCnn / l 7P7 / B / Y antibodies can be "repurposed" by converting them into antibodies with different binding properties, and therefore different functions, through somatic mutation. Antibodies can have one of the ideal folds and structures for evolving binding specificity through limited mutation, which should impact the vast capacity of the natural immune response to recognize essentially infinite varieties of foreign antigens. Furthermore, this work shows, for the first time, a general modification pathway for evolving a dual-specific antibody from any single-specific antibody, summarized below. Using mutagenesis analyses (e.g., alanine scan), the region of the antigen-binding site that tolerates mutation without severely altering the binding of its primary antigen can first be identified. Other modalities All patents, patent applications, patent application publications and other publications indicated or referenced in this descriptive memorandum are incorporated herein by this reference to the same extent as if each patent, patent application, patent application publication or separate publication were specifically and individually indicated as incorporated herein by this reference.

Claims

1. A method for making a specific double antibody or antigen-binding fragment thereof, comprising a variable heavy chain domain (Vh) and a variable light chain domain (Vl), wherein the Vh and Vl of the specific double antibody are paired to form an antigen-binding site that binds specifically to a first epitope and a second epitope, wherein said method comprises the steps of: (a) providing an antibody with a Vh and Vl, wherein the Vh and Vl are paired to form an antigen-binding site that binds to a first epitope but not to the second epitope and wherein said antibody comprises at least one amino acid at position 32, 50, or 91 of the Vl that is electrostatic or hydrophobic, (b) altering the nucleic acid sequence encoding the Vh of the antibody of step (a), wherein one or more solvent-accessible amino acid residues are altered, (c) expressing the altered Vl and Vh of step (b),and (d) selecting a specific double antibody or antigen-binding fragment thereof, comprising the Vl and the altered Vh of step (c), wherein the Vh and Vl are paired to form an antigen-binding site that binds specifically to the first epitope and the second epitope.

2. The method of claim 1, wherein at least two of the amino acids at position 32, 50 or 91 are electrostatic or hydrophobic.

3. The method of claim 1, wherein the three amino acids at positions 32, 50 and 91 are electrostatic or hydrophobic.

4. The method of any of claims 1-3, wherein the electrostatic residue is a tyrosine.

5. The method of any of claims 1-3, wherein the hydrophobic residue is a tryptophan.

6. The method of any of claims 1-5, wherein the nucleic acid sequence encoding Vh is altered according to the diversity of multiple naturally occurring heavy chain amino acid sequences.

7. The method of any of claims 1-6, wherein the solvent-exposed residue position is an amino acid residue position selected from the group consisting of positions 33, 34, 50-58 and 95-97 of Vh.

8. The method of any of claims 1-7 further comprising altering the nucleic acid sequence encoding the V1 of the antibody of step (a), wherein one or more solvent-accessible amino acid residues are altered. Rbccnn / ι ζηζ / E / γ 9. The method of claim 8, wherein the solvent-exposed residue position is an amino acid residue position selected from amino acids 93-96 of VI.

10. The method of any of claims 1-9, wherein the altered Vh are expressed in phages with the Vl during the selection of step (d).

11. The method of any of claims 1-10, wherein the antibody of step (a) comprises a light chain variable region complementarity-determining region CDRL1 comprising the amino acid sequence KASQSVINDAA (SEQ ID NO: 9), a CDRL2 comprising the amino acid sequence YTSHRYT (SEQ ID NO: 10), and a CDRL3 comprising the amino acid sequence QQDYTSPWTF (SEQ ID NO: 11).

12. The method of any of claims 1-11, wherein the antibody of step (a) comprises a heavy chain variable region complementarity-determining region CDRH1 comprising the amino acid sequence DYSMH (SEQ ID NO: 13), a CDRH2 comprising the amino acid sequence VWINTETGEPTYADDFK (SEQ ID NO: 17), and a CDRH3 comprising the amino acid sequence GGIFYGMDY (SEQ ID NO: 20).

13. The method of any of claims 1-12, wherein the antigen-binding site of the double-specific antibody of step (d) binds to the first epitope and the second epitope in a mutually exclusive manner.

14. The method of any of claims 1-12, wherein the antigen-binding site of the double-specific antibody of step (d) binds to the first epitope and the second epitope simultaneously.

15. The method of any of claims 1-14, wherein the first epitope is derived from a biological molecule and the second epitope is derived from the same biological molecule.

16. The method of any of claims 1-14, wherein the first epitope is derived from a first biological molecule and the second epitope is derived from a second biological molecule.

17. The method of claim 16, wherein the first biological molecule and the second biological molecule are selected from the group consisting of IL4 / IL5, IL4 / IL13.

18. The method of claim 16, wherein the first biological molecule and the second biological molecule are cytokines. RbCCnn / l 7Π7 / Β / Y 19. The method of claim 16, wherein the first or second biological molecule is a molecule that can increase the half-life of the specific double antibody when it binds to the antibody in vivo.

20. The method of claim 16, wherein the first or second biological molecule is serum albumin or a neonatal Fe receptor (FcRn).

21. The method of claim 16, wherein the first or second biological molecule is a molecule that can enhance the effector function of a specific double antibody when bound to the antibody in vivo.

22. The method of claim 16, wherein the first or second biological molecule is attached to a protein on the cell surface in natural cytolytic cells or macrophages.

23. The method of claim 22, wherein said cell surface protein is a receptor for Fe or C1q.

24. The method of any of claims 1-23, wherein the Vh and Vl of the double specific antibody are paired to form an antigen-binding site that binds specifically to the first epitope or the second epitope with a Kd of 10⁶ or less.

25. The method of claim 24, wherein the Vh and Vl of the double specific antibody are paired to form an antigen-binding site that binds specifically to the first epitope or the second epitope with a Kd of 10-9 or less.

26. The method of claim 25, wherein the Vh and Vl of the double specific antibody are paired to form an antigen-binding site that binds specifically to the first epitope or the second epitope with a Kd of 10'12 or less.

27. The method of any of claims 1-23, wherein the Vh and Vl of the double specific antibody are paired to form an antigen-binding site that binds specifically to the first epitope and the second epitope with a Kd of 10⁶ or less.

28. The method of claim 27, wherein the Vh and Vl of the specific double antibody are paired to form an antigen-binding site that binds specifically to the first epitope and the second epitope with a Kode 10 9o smaller. RbCCnn / l 7P7 / B / Y 29. The method of claim 28, wherein the Vh and Vl of the double specific antibody are paired to form an antigen-binding site that binds specifically to the first epitope and the second epitope with a Kd of 1012 or less.

30. The method of any of claims 1-29, wherein the first biological molecule and the second biological molecule are not structurally similar.

31. The method of any of claims 1-30, wherein the selection of step (d) comprises deep sequencing.

32. A specific double isolated antibody or antigen-binding fragment thereof manufactured by the process of claim 1.

33. The isolated double specific antibody of claim 32, wherein the double specific antibody is a monoclonal antibody.

34. The isolated double specific antibody of claim 32, wherein the fragment is a Fab or an scFv.

35. The isolated double specific antibody of claim 32, wherein the double specific antibody is an IgG.

36. An isolated double-specific antibody or antigen-binding fragment thereof comprising the amino acid sequence of any of the antibodies in Figures 4A, 4B, 4C, 7A, 7C, or 7D.

37. A specific double antibody isolated or antigen-binding fragment thereof comprising the six CDRs below: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10); (iii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11); (iv) a CDRH1 comprising the amino acid sequence of DYDIH (SEQ ID NO: 14); (v) a CDRH2 comprising the amino acid sequence of VWINTETGEPTYADDFK (SEQ ID NO: 17); and (vi) a CDRH3 comprising the amino acid sequence of EILFYGMDY (SEQ ID NO: 21).

38. A specific double antibody isolated or antigen-binding fragment thereof comprising the six CDRs below: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10); (iii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11); (iv) a CDRH1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15); (v) a CDRH2 comprising the amino acid sequence of AGIVYDATGFTTYADDFK (SEQ ID NO: 18); and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20).

39. A specific double antibody isolated or antigen-binding fragment thereof comprising the six CDRs below: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10); (ii) a CDRL3 comprising the amino acid sequence of QQDYTPFPLTF (SEQ ID NO: 12); (iv) a CDRH1 comprising the amino acid sequence of DYLMH (SEQ ID NO: 16); (v) a CDRH2 comprising the amino acid sequence of AVIVSITGRTYYADDFK (SEQ ID NO: 19); and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20).

40. A specific double antibody isolated or antigen-binding fragment thereof comprising the six CDRs below: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10); (ii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11); (iv) a CDRH1 comprising the amino acid sequence of DYSMH (SEQ ID NO: 13); (v) a CDRH2 comprising the amino acid sequence of GVIFQSGATYYADDFK (SEQ ID NO: 22); and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20).

41. A specific double antibody isolated or antigen-binding fragment thereof comprising the six CDRs below: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10); (iii) a CDRL3 comprising the amino acid sequence of QQDYTSPWTF (SEQ ID NO: 11); (iv) a CDRH1 comprising the amino acid sequence of DYSMH (SEQ ID NO: 13); (v) a CDRH2 comprising the amino acid sequence of GVIFQSGATYYADDFK (SEQ ID NO: 23); and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20).

42. A specific double antibody isolated or antigen-binding fragment thereof comprising the six CDRs below: RbCCnn / l 7P7 / B / Y (i) a CDRL1 comprising the amino acid sequence KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence YTSHRYT (SEQ ID NO: 10); (iii) a CDRL3 comprising the amino acid sequence QQDYX1X2PWTF (SEQ ID NO: 24), wherein X1 is Thr, Ie, Leu or Lys, and X2 is Ser or His; (iv) a CDRH1 comprising the amino acid sequence DYFIH (SEQ ID NO: 15); (v) a CDRH2 comprising the amino acid sequence X1GIVYDATGFTX2YA X3X4FK (SEQ ID NO: 25), where X1 is Ala or Gly, X2 is Thr, Lie, Val or Ala, X3 is Asp, Val or Glu, and X4 is Asp, Glu, Asn, Ser, Lie, Leu, Thr, Ala or Phe; and (vi) a CDRH3 comprising the amino acid sequence GGIFYGMDY (SEQ ID NO: 20).

43. The isolated double specific antibody or antigen-binding fragment thereof of claim 41 comprising the six CDRs below: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10); (iii) a CDRL3 comprising the amino acid sequence of QQDYTHPWTF (SEQ ID NO: 27); (iv) a CDRH1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15); (v) a CDRH2 comprising the amino acid sequence of GGIVYDATGFTTYAEEFK (SEQ ID NO: 28); and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20).

44. The isolated double specific antibody or antigen-binding fragment thereof of claim 41 comprising the six CDRs below: (i) a CDRL1 comprising the amino acid sequence of KASQSVINDAA (SEQ ID NO: 9); (ii) a CDRL2 comprising the amino acid sequence of YTSHRYT (SEQ ID NO: 10); (iii) a CDRL3 comprising the amino acid sequence of QQDYKHPWTF (SEQ ID NO: 31); (iv) a CDRH1 comprising the amino acid sequence of DYFIH (SEQ ID NO: 15); (v) a CDRH2 comprising the amino acid sequence of AGIVYDATGFTVYADDFK (SEQ ID NO: 32); and (vi) a CDRH3 comprising the amino acid sequence of GGIFYGMDY (SEQ ID NO: 20).

45. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 42-44, further comprising a frame region 3 (FR3) comprising the amino acid sequence GRX1TITX2DX3STSTX4 (SEQ ID NO: 26), wherein X1 is Val or Phe, X2 is Arg or lie, X3 is Thr, Phe, Met or Pro, and X4 is Ala or Val.

46. ​​A specific double antibody isolated or antigen-binding fragment thereof comprising a light chain variable region selected from the amino acid sequence of SEQ ID NO: 1, 5, 29 or 33 and a heavy chain variable region selected from SEQ ID NO: 2, 3, 4, 6, 7, 8, 30 or 34. RbCCnn / l 7P7 / B / Y 47. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 38, 39 and 42-46, wherein said antibody or antigen-binding fragment thereof binds to IL4 with a Kd of 500 nM or less and IL5 with a Kd of about 900 nM or less.

48. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 38, 39 and 42-46, wherein said antibody binds to IL4 with a Kd of 100 nM or less and to IL5 with a Kd of about 100 nM or less.

49. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 38, 39 and 42-46, wherein said antibody binds to IL4 with a Kd of 10 nM or less and to IL5 with a Kd of about 50 nM or less.

50. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 40, 41 and 46, wherein said antibody binds to IL4 with a Kd of 500 nM or less and to IL13 with a Kd of about 900 nM or less.

51. The isolated double specific antibody of any of claims 40, 41 and 46, wherein said antibody binds to IL4 with a Kd of 100 nM or less and to IL13 with a Kd of about 100 nM or less.

52. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 36-51, wherein said antibody inhibits or blocks the binding of IL4, IL5 or IL13 to its receptor.

53. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 36-51, wherein said antibody is a monoclonal antibody.

54. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 36-51, wherein said antibody is an IgG antibody.

55. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 36-51, wherein said fragment is a Fab fragment or a single-chain variable fragment (scFv).

56. The isolated double-specific antibody or antigen-binding fragment thereof of any of claims 36-51, wherein at least a portion of the framework sequence is a human consensus framework sequence. RbCCnn / l 7P7 / B / Y 57. The isolated double specific antibody or antigen-binding fragment thereof of any of claims 32-51, wherein the antibody is a chimeric, humanized, or fully human antibody.

58. A pharmaceutical composition comprising any of the antibodies of claims 32-57.

59. A polynucleotide encoding a specific double antibody isolated or antigen-binding fragment thereof of any of claims 32-57.

60. A vector comprising the polynucleotide of claim 59.

61. A host cell comprising the vector of claim 60.

62. A method for producing the antibody or antigen-binding fragment thereof of any of claims 32-57, wherein said method comprises culturing a host cell comprising a vector of claim 60 and recovering said antibody.

63. A method for treating asthma in a subject, wherein said method comprises administering to said subject the antibody or antibody fragment thereof of any of claims 32-57, wherein said administration occurs for a time and in a quantity sufficient to treat or prevent said asthma in said subject.

64. The method of claim 61, wherein said method also comprises administering at least one additional asthma treatment selected from the group consisting of an IgE antagonist, an antihistamine, threophylline, salbutamol, beclomethasone dipropionate, sodium cromoglycate, a steroid, and anti-inflammatory agents.

65. The method of claim 63 or 64, wherein the asthma is allergic asthma.

66. A method for treating a proliferative disorder in a subject, wherein said method comprises administering to said subject the antibody or antibody fragment thereof or any of claims 32-57, wherein said administration occurs for a time and in a quantity sufficient to treat said proliferative disorder in said subject.

67. The method of claim 66, wherein said proliferative disorder is cancer.

68. The method of claim 67, further comprising administering to said subject an additional antiproliferative agent selected from the group consisting of a chemotherapeutic agent, a cytotoxic agent, and an antiangiogenic agent.