Binding constructs and methods for use thereof

Abstract

The invention relates to novel binding domain-immunoglobulin fusion proteins that feature a binding domain for a cognate structure such as an antigen, a counterreceptor or the like, a wild-type IgG1, IGA or IgE hinge-acting region, i.e., IgE CH2, region polypeptide or a mutant IgG1 hinge region polypeptide having either zero, one or two cysteine residues, and immunoglobulin CH2 and CH3 domains, and that are capable of ADCC and/or CDC while occurring predominantly as polypeptides that are compromised in their ability to form disulfide-linked multimers. The fusion proteins can be recombinantly produced at high expression levels. Also provided are related compositions and methods, including cell surface forms of the fusion proteins and immunotherapeutic applications of the fusion proteins and of polynucleotides encoding such fusion proteins.

1 .- 413 . (canceled) 
     
     
         414 . A method of treating a B-cell disorder, comprising administering to a patient a therapeutically effective amount of a fusion protein that binds CD20 and comprises an amino acid sequence as set forth in:
 SEQ ID NO:135 (2H7 scFv (CSS—S)H WCH2 WCH3), wherein proline at position 283 is substituted with serine;   SEQ ID NO:137 (2H7 scFv (SCS—S)H WCH2 WCH3), wherein proline at position 283 is substituted with serine;   SEQ ID NO:166 (2H7 scFv (CSC—S)H WCH2 WCH3), wherein proline at position 283 is substituted with serine;   SEQ ID NO:372 (2H7 scFv VHL11S(CSS—S)H WCH2 WCH3);   SEQ ID NO:246 (2H7 scFv VH L11S(CSC—S)H WCH2 WCH3);   SEQ ID NO:370 (2H7 scFv VH L11S(SSS—S)H WCH2 WCH3);   SEQ ID NO:268 (2H7 scFv VH L11S(CSS—S)H K322S CH2 WCH3); or   SEQ ID NO:276 (2H7 scFv VH L11S(CSS—S)H P331S CH2 WCH3).   
     
     
         415 . The method of  claim 414 , wherein the B-cell disorder is a B-cell lymphoma or chronic lymphocytic leukemia. 
     
     
         416 . The method of  claim 414 , wherein the B-cell disorder is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, type I diabetes mellitus, multiple sclerosis, immune thrombocytopenic purpura, psoriasis, inflammatory bowel disease, Crohn's disease, and ulcerative colitis. 
     
     
         417 . A method of treating a B-cell disorder, comprising administering to a patient a therapeutically effective amount of a fusion protein that binds CD20 and consists essentially of an amino acid sequence as set forth in SEQ ID NO:135 (2H7 scFv (CSS—S)H WCH2 WCH3) wherein proline at position 283 is substituted with serine, SEQ ID NO:166 (2H7 scFv (CSC—S)H WCH2 WCH3) wherein proline at position 283 is substituted with serine, SEQ ID NO:372 (2H7 scFv VHL11S(CSS—S)H WCH2 WCH3), or SEQ ID NO:246 (2H7 scFv VH L11S(CSC—S)H WCH2 WCH3). 
     
     
         418 . The method of  claim 417 , wherein the B-cell disorder is a B-cell lymphoma or chronic lymphocytic leukemia. 
     
     
         419 . The method of  claim 417 , wherein the B-cell disorder is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, type I diabetes mellitus, multiple sclerosis, immune thrombocytopenic purpura, psoriasis, inflammatory bowel disease, Crohn's disease, and ulcerative colitis. 
     
     
         420 . A method of treating a B-cell disorder, comprising administering to a patient a therapeutically effective amount of a fusion protein that comprises from amino-terminus to carboxy-terminus: (i) an scFv binding domain polypeptide that binds CD20, wherein the scFv comprises amino acids 23-265 of SEQ ID NO:246; (ii) an immunoglobulin hinge polypeptide consisting of amino acids 269-283 as set forth in SEQ ID NO:246; and (iii) an amino-terminally truncated immunoglobulin heavy chain constant region polypeptide consisting of amino acids 284-500 as set forth in SEQ ID NO:246. 
     
     
         421 . The method of  claim 420  wherein the B-cell disorder is a B-cell lymphoma or chronic lymphocytic leukemia. 
     
     
         422 . The method of  claim 420  wherein the B-cell disorder is selected from the group consisting of rheumatoid arthritis, systemic lupus erythematosus, type I diabetes mellitus, multiple sclerosis, immune thrombocytopenic purpura, psoriasis, inflammatory bowel disease, Crohn's disease, and ulcerative colitis.