Multivalent antibodies and uses therefor

Abstract

The present application describes engineered antibodies, with three or more functional antigen binding sites, and uses, such as therapeutic applications, for such engineered antibodies.

What is claimed is:  
     
         1 . An isolated antibody comprising an Fc region and three or more antigen binding sites amino-terminal to the Fc region.  
     
     
         2 . The antibody of  claim 1  comprising four antigen binding sites.  
     
     
         3 . The antibody of  claim 1  comprising five or more antigen binding sites.  
     
     
         4 . The antibody of  claim 1  comprising a polypeptide chain, wherein the polypeptide chain comprises two or more variable domains.  
     
     
         5 . The antibody of  claim 4  wherein the polypeptide chain comprises VD1-(X1) n -VD2-(X2) n -Fc, wherein VD1 is a first variable domain, VD2 is a second variable domain, Fc is one polypeptide chain of an Fc region, X1 and X2 represent an amino acid or polypeptide, and n is 0 or 1.  
     
     
         6 . The antibody of  claim 5  comprising two or more polypeptide chains, each comprising VD1-(X1) n -VD2-(X2) n -Fc.  
     
     
         7 . The antibody of  claim 1  comprising at least one polypeptide chain with the formula: 
 (a) VH-CH1-flexible linker-VH-CH1-Fc region chain; or  
 (b) VH-CH1-VH-CH1-Fc region chain.  
 
     
     
         8 . The antibody of  claim 1  comprising at least two light chain variable domain polypeptides.  
     
     
         9 . The antibody of  claim 8  wherein the light chain variable domain polypeptides further comprise a CL domain.  
     
     
         10 . The antibody of  claim 1  comprising a polypeptide chain, wherein the polypeptide chain comprises Fd-flexible linker-Fd.  
     
     
         11 . The antibody of  claim 10  wherein the flexible linker comprises a peptide selected from the group consisting of gly-ser, gly-ser-gly-ser (SEQ ID NO:10), ala-ser, and gly-gly-gly-ser (SEQ ID NO:11).  
     
     
         12 . The antibody of  claim 1  which is internalized faster than a bivalent antibody by a cell expressing an antigen to which the antibodies bind.  
     
     
         13 . The antibody of  claim 1  which is an agonist antibody.  
     
     
         14 . The antibody of  claim 1  which induces apoptosis.  
     
     
         15 . The antibody of  claim 1  wherein the three or more antigen binding sites all bind the same antigen.  
     
     
         16 . The antibody of  claim 1  wherein the three or more antigen binding sites bind two or more different antigens.  
     
     
         17 . The antibody of  claim 1  which binds a cell surface protein expressed by tumor cells.  
     
     
         18 . The antibody of  claim 17  wherein the cell surface protein is selected from the group consisting of the Epidermal Growth Factor Receptor (EGFR), HER2 receptor, HER3 receptor, HER4 receptor and DcR3 receptor.  
     
     
         19 . The antibody of  claim 17  wherein the cell surface protein is HER2 receptor.  
     
     
         20 . The antibody of  claim 1  which binds a cell surface protein overexpressed by tumor cells.  
     
     
         21 . The antibody of  claim 1  which binds a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily.  
     
     
         22 . The antibody of  claim 21  wherein the TNF receptor is an Apo2L receptor.  
     
     
         23 . The antibody of  claim 22  wherein the Apo2L receptor is selected from the group consisting of DR4, DR5, DcR1 and DcR2.  
     
     
         24 . The antibody of  claim 22  wherein the Apo2L receptor is DR4 or DR5.  
     
     
         25 . The antibody of  claim 21  which is an agonist antibody.  
     
     
         26 . The antibody of  claim 21  which induces apoptosis.  
     
     
         27 . The antibody of  claim 1  which binds a B cell surface antigen.  
     
     
         28 . The antibody of  claim 27  wherein the B cell surface antigen is selected from the group consisting of CD19, CD20, CD22 and CD40.  
     
     
         29 . The antibody of  claim 27  wherein the B cell surface antigen is CD20.  
     
     
         30 . An immunoconjugate comprising the antibody of  claim 1  conjugated with a cytotoxic agent.  
     
     
         31 . The immunoconjugate of  claim 30  wherein the cytotoxic agent is active in killing cells once internalized.  
     
     
         32 . The immunoconjuate of  claim 30  wherein the cytotoxic agent is selected from the group consisting of a radioactive isotope, a maytansinoid and a calecheamicin.  
     
     
         33 . An isolated antibody comprising three or more antigen binding sites, wherein the antibody is capable of binding a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily.  
     
     
         34 . The antibody of  claim 33  which is not a native sequence IgM or IgA antibody.  
     
     
         35 . The antibody of  claim 33  which has only one Fc region or lacks an Fc region.  
     
     
         36 . The antibody of  claim 33  which comprises a polypeptide chain, wherein the polypeptide chain comprises two or more variable domains.  
     
     
         37 . The antibody of  claim 33  which comprises four antigen binding sites each capable of binding the TNF receptor.  
     
     
         38 . The antibody of  claim 33  wherein the TNF receptor is an Apo2L receptor.  
     
     
         39 . The antibody of  claim 38  wherein the Apo2L receptor is selected from the group consisting of DR4, DR5, DcR1 and DcR2.  
     
     
         40 . The antibody of  claim 38  wherein the Apo2L receptor is DR4 or DR5.  
     
     
         41 . The antibody of  claim 33  which is an agonist antibody.  
     
     
         42 . The antibody of  claim 33  which induces apoptosis.  
     
     
         43 . An isolated antibody comprising at least three antigen binding sites, wherein the antibody is capable of binding an ErbB receptor.  
     
     
         44 . The antibody of  claim 43  which is not a native sequence IgM or IgA antibody.  
     
     
         45 . The antibody of  claim 43  which has only one Fc region or lacks an Fc region.  
     
     
         46 . The antibody of  claim 43  which comprises a polypeptide chain, wherein the polypeptide chain comprises two or more variable domains.  
     
     
         47 . The antibody of  claim 43  which comprises four antigen binding sites each capable of binding the ErbB receptor.  
     
     
         48 . The antibody of  claim 43  wherein the ErbB receptor is selected from the group consisting of Epidermal Growth Factor Receptor (EGFR), HER2 receptor, HER3 receptor and HER4 receptor.  
     
     
         49 . The antibody of  claim 43  where the ErbB receptor is HER2 receptor.  
     
     
         50 . An isolated antibody comprising at least three antigen binding sites, wherein the antibody is capable of binding a B cell surface antigen.  
     
     
         51 . The antibody of  claim 50  which is not a native sequence IgM or IgA antibody.  
     
     
         52 . The antibody of  claim 50  which has only one Fc region or lacks an Fc region.  
     
     
         53 . The antibody of  claim 50  which comprises a polypeptide chain, wherein the polypeptide chain comprises two or more variable domains.  
     
     
         54 . The antibody of  claim 50  which comprises four antigen binding sites each capable of the B cell surface antigen.  
     
     
         55 . The antibody of  claim 50  wherein the B cell surface antigen is selected from the group consisting of CD19, CD20, CD22 and CD40.  
     
     
         56 . An isolated antibody comprising at least three antigen binding sites, wherein the antibody is capable of binding an antigen which is overexpressed by cancer cells.  
     
     
         57 . A polypeptide chain comprising: 
 (a) VH-CH1-flexible linker-VH-CH1-dimerization domain; or    (b) VH-CH1-VH-CH1-dimerization domain.    
     
     
         58 . An isolated antibody comprising the polypeptide chain of  claim 57 .  
     
     
         59 . The antibody of  claim 58  further comprising two or more light chain variable domain polypeptides.  
     
     
         60 . The antibody of  claim 59  wherein the light chain variable domain polypeptides comprise VL-CL.  
     
     
         61 . An isolated antibody comprising a dimerization domain and three or more antigen binding sites amino-terminal thereto  
     
     
         62 . The antibody of  claim 61  wherein the dimerization domain is selected from the group consisting of a hinge region, an Fc region, a CH3 domain, and a CH4 domain.  
     
     
         63 . The antibody of  claim 62  wherein the dimerization domain is a hinge region.  
     
     
         64 . The antibody of  claim 63  wherein the dimerization domain further comprises a leucine zipper.  
     
     
         65 . The antibody of  claim 63  comprising a polypeptide chain comprising the formula: 
 (a) VH-CH1-flexible linker-VH-CH1-hinge region; or  
 (b) VH-CH1-VH-CH1-hinge region.  
 
     
     
         66 . A polypeptide chain comprising three or more heavy chain or light chain variable domains, wherein each of the variable domains is able to combine with three or more light chain or heavy chain variable domain polypeptides to form three or more antigen binding sites, each directed against the same antigen.  
     
     
         67 . The polypeptide chain of  claim 66  which comprises three heavy chain variable domains which are able to combine with three light chain variable domain polypeptides to form three antigen binding sites directed against the same antigen.  
     
     
         68 . The polypeptide chain of  claim 66  which comprises four heavy chain variable domains which are able to combine with four light chain variable domain polypeptides to form four antigen binding sites directed against the same antigen.  
     
     
         69 . The polypeptide chain of  claim 66  wherein the antigen is a receptor in the Tumor Necrosis Factor (TNF) receptor superfamily.  
     
     
         70 . The polypeptide chain of  claim 66  wherein the antigen is a B cell surface antigen.  
     
     
         71 . The polypeptide chain of  claim 66  wherein the antigen is an ErbB receptor.  
     
     
         72 . The polypeptide chain of  claim 66  wherein the antigen is a cell surface protein expressed by tumor cells.  
     
     
         73 . A polypeptide chain comprising the formula: 
 (a) VL-CL-flexible linker-VL-CL-flexible linker-VL-CL;    (b) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1;    (c) (VL-CL) n , wherein n is three or more; or    (d) (VH-CH1) n , wherein n is three or more.    
     
     
         74 . The polypeptide chain of  claim 67  comprising the formula: 
 (a) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1;  
 (b) VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1-flexible linker-VH-CH1; or  
 (c) (VH-CH1) n , wherein n is three or four.  
 
     
     
         75 . An isolated antibody comprising the polypeptide chain of  claim 66 .  
     
     
         76 . The isolated antibody of  claim 75  further comprising the three or more light chain or heavy chain variable domain polypeptides.  
     
     
         77 . The isolated antibody of  claim 76  comprising three or more light chain variable domain polypeptides, each comprising VL-CL.  
     
     
         78 . The isolated antibody of  claim 77  comprising four light chain variable domain polypeptides, each comprising VL-CL.  
     
     
         79 . A polypeptide chain comprising: 
 (a) VL-CL-flexible linker-VL-CL-dimerization domain;    (b) VL-CL-VL-CL-dimerization domain.    
     
     
         80 . An immunoconjugate comprising the antibody of  claim 75  conjugated with a cytotoxic agent.  
     
     
         81 . Isolated nucleic acid encoding the antibody of  claim 1 , the polypeptide chain of  claim 57  or the polypeptide chain of  claim 66 .  
     
     
         82 . A vector comprising the nucleic acid of  claim 81 .  
     
     
         83 . A host cell comprising the nucleic acid of  claim 81 .  
     
     
         84 . A process of producing an antibody or polypeptide chain comprising culturing the host cell of  claim 83  so that the nucleic acid is expressed.  
     
     
         85 . The process of  claim 84  further comprising recovering the antibody or polypeptide chain from the host cell culture.  
     
     
         86 . The process of  claim 85  wherein the antibody or polypeptide chain is recovered from the host cell culture medium.  
     
     
         87 . A method for treating a disorder in a mammal comprising administering to the mammal a therapeutically effective amount of the antibody of  claim 1 .  
     
     
         88 . The method of  claim 87  wherein the disorder is cancer.  
     
     
         89 . The method of  claim 87  further comprising administering a therapeutically effective amount of a cytotoxic agent to the mammal.  
     
     
         90 . A method of inducing apoptosis of a cancer cell comprising exposing the cell to the antibody of  claim 33 .  
     
     
         91 . A method of killing a B cell comprising exposing the B cell to the antibody of  claim 50 .  
     
     
         92 . A method of killing a cell which expresses an ErbB receptor comprising exposing the cell to the antibody of  claim 43 .  
     
     
         93 . The method of  claim 92  wherein the cell is a cancer cell which overexpresses an ErbB receptor.