Dual specificity antibodies and methods of making and using

Abstract

Antibodies having dual specificity for two different but structurally related antigens are provided. The antibodies can be, for example, entirely human antibodies, recombinant antibodies, or monoclonal antibodies. Preferred antibodies have dual specificity for IL-1α and IL-1β and neutralize IL-1α and IL-1β activity in vitro and in vivo. An antibody of the invention can be a full-length antibody or an antigen-binding portion thereof. Methods of making and methods of using the antibodies of the invention are also provided. The antibodies, or antibody portions, of the invention are useful for detecting two different but structurally related antigens (e.g., IL-1α and IL-1β) and for inhibiting the activity of the antigens, e.g., in a human subject suffering from a disorder in which IL-1α and/or IL-1β activity is detrimental.

1 - 69 . (canceled) 
     
     
         70 . A method of making an antibody or an antigen binding portion thereof library comprising the steps of:
 a) obtaining a recombinant heavy chain or an antigen binding portion thereof library A from an antibody repertoire resulting from exposure to a first antigen;   b) obtaining a recombinant light chain or an antigen binding portion thereof library B from an antibody repertoire resulting from exposure to the first antigen;   c) obtaining a recombinant heavy chain or an antigen binding portion thereof library C from an antibody repertoire resulting from exposure to a second antigen;   d) obtaining a recombinant light chain or an antigen binding portion thereof library D from an antibody repertoire resulting from exposure to the second antigen; and   e) combining the recombinant heavy chain or an antigen binding portion thereof library A with the recombinant light chain or an antigen binding portion thereof library D to obtain an antibody or an antigen binding portion thereof library X and/or combining the recombinant heavy chain or an antigen binding portion thereof library C with the recombinant light chain or an antigen binding portion thereof library to obtain an antibody or an antigen binding portion thereof library Y.   
     
     
         71 . A method of making an antibody or an antigen binding portion thereof library according to  claim 70  further comprising the step of combining the antibody or an antigen binding portion thereof library X with the antibody or an antigen binding portion thereof library Y to obtain an antibody or an antigen binding portion thereof library Z. 
     
     
         72 . The antibody or an antigen binding portion thereof library X made according to the method of  claim 70 . 
     
     
         73 . The antibody or an antigen binding portion thereof library Y made according to the method of  claim 70 . 
     
     
         74 . The antibody or an antigen binding portion thereof library Z made according to the method of  claim 71 . 
     
     
         75 . A method of making a dual specific antibody or an antigen binding portion thereof comprising the steps of:
 a) obtaining a recombinant heavy chain or an antigen binding portion thereof library A from an antibody repertoire resulting from exposure to a first antigen;   b) obtaining a recombinant light chain or an antigen binding portion thereof library B from an antibody repertoire resulting from exposure to the first antigen;   c) obtaining a recombinant heavy chain or an antigen binding portion thereof library C from a antibody repertoire resulting from exposure to a second antigen;   d) obtaining a recombinant light chain or an antigen binding portion thereof library D from an antibody repertoire resulting from exposure to the second antigen;   e) combining the recombinant heavy chain or an antigen binding portion thereof library A with the recombinant light chain or an antigen binding portion thereof library D to obtain an antibody or an antigen binding portion thereof library X and/or combining the recombinant heavy chain or an antigen binding portion thereof library C with the recombinant light chain or an antigen binding portion thereof library B to obtain an antibody or an antigen binding portion thereof library Y; and   f) selecting from the antibody or an antigen binding portion thereof library X and/or antibody or an antigen binding portion thereof library Y an antibody or an antigen binding portion thereof that binds both the first and the second antigen.   
     
     
         76 . The dual specific antibody made by the method of  claim 75 . 
     
     
         77 . A method of making a dual specific antibody or an antigen binding portion thereof comprising the steps of:
 a) obtaining a recombinant heavy chain or an antigen binding portion thereof library A from an antibody repertoire resulting from exposure to a first antigen;   b) obtaining a recombinant light chain or an antigen binding portion thereof library B from an antibody repertoire resulting from exposure to the first antigen;   c) obtaining a recombinant heavy chain or an antigen binding portion thereof library C from an antibody repertoire resulting from exposure to a second antigen;   d) obtaining a recombinant light chain or an antigen binding portion thereof library D from an antibody repertoire resulting from exposure to the second antigen;   e) combining the recombinant heavy chain or an antigen binding portion thereof library A with the recombinant light chain or an antigen binding portion thereof library D to obtain an antibody library X and/or combining the recombinant heavy chain or an antigen binding portion thereof library C with the recombinant light chain or an antigen binding portion thereof library B to obtain an antibody or an antigen binding portion thereof library Y;   f) combining the antibody or an antigen binding portion thereof library X with the antibody library or an antigen binding portion thereof Y to obtain an antibody or an antigen binding portion thereof library Z; and   g) selecting from the antibody or an antigen binding portion thereof library Zan antibody or an antigen binding portion thereof that binds both the first and the second antigen.   
     
     
         78 . The dual specific antibody or an antigen binding portion thereof made by the method according to  claim 77 . 
     
     
         79 . The method according to  claim 70  wherein the first and second antigen is each independently selected from the group consisting of proteins, polypeptides and peptides provided that the first and second antigens are not the same. 
     
     
         80 . The method of  claim 79 , wherein the proteins, polypeptides and peptides are secreted proteins or surface receptors. 
     
     
         81 . The method of  claim 80 , wherein the secreted protein is selected from the group consisting of an MN, a TNF, an interleukin, PF4, a CRO, 9E3, EMAP-II, a CSF, an FGF, and a PDGF. 
     
     
         82 . The method according to  claim 81 , therein the first antigen is IL-1α and the second antigen is IL-1β. 
     
     
         83 - 88 . (canceled) 
     
     
         89 . A method of making a dual specific antibody or an antigen binding portion thereof, the method comprising:
 a) obtaining a first recombinant heavy chain or an antigen binding portion thereof library by exposure of an antibody repertoire to a first antigen;   b) obtaining a second recombinant light chain or an antigen binding portion thereof library by exposure of an antibody repertoire to a first antigen;   c) obtaining a third recombinant heavy chain or an antigen binding portion thereof library by exposure of an antibody repertoire to a second antigen;   d) obtaining a forth recombinant light chain or an antigen binding portion thereof library by exposure of an antibody repertoire to a second antigen;   e) combining the first library with the forth library to obtain a fifth antibody or an antigen binding portion thereof library and/or combining the second library with the third library to obtain a sixth antibody or an antigen binding portion thereof library; and identifying from the fifth or sixth library an antibody or an antigen binding portion thereof that specifically binds to the first and second antigens.   
     
     
         90 . The method of  claim 89 , wherein the first, second, third or fourth libraries comprise human antibody chains or antigen binding portions thereof. 
     
     
         91 . The method of  claim 89 , wherein the first, second, third and fourth libraries comprise human antibody chains or antigen binding portions thereof. 
     
     
         92 . The method of  claim 89 , wherein the first and second antigens are structurally related antigens. 
     
     
         93 . The method of  claim 89 , wherein the first and second antigens are each independently selected from the group consisting of caspases, cytokines, secreted proteinases, and cytokine receptor families, TNF receptor family members, TGFβ receptor family members, EGF receptor family members, FGF receptor family members, PDGF receptor family members, VEGF receptor family members and angiopoietin receptor family members. 
     
     
         94 . The method of  claim 93 , wherein the cytokines are selected from the group consisting of TNF family members IL-6 family members, interferons, TGFβ family members, EGF family members, FGF family members, PDGF family members, VEGF family members, angiopoietins, bone morphogenetic proteins. 
     
     
         95 . The method of  claim 89 , wherein the first and second antigens are each independently selected from the group consisting of IP-10, PF4, a GRO, 9E3, EMAP-II, a CSF, an FGF, and a PDGF. 
     
     
         96 . The method of  claim 89 , wherein the first antigen and the second antigen are IL-1α and IL-1β; IL-1α and IL-18; IL-1β and IL-18; EGFR and IGFR; HER2 and HER3; and EGFR and HER3, respectively.