Deimmunized Serum-Binding Domains and Their Use in Extending Serum Half-Life

Abstract

The present invention is directed to a polypeptide (for example, an antigen-binding molecule) that comprises a polypeptide portion of a deimmunized serum-binding protein capable of binding to said serum protein. The presence of the serum-binding protein extends the serum half-life of the polypeptide, relative to the serum half-life of the polypeptide if lacking the polypeptide portion of the deimmunized serum-binding protein. The invention also pertains to methods and uses that employ such molecules.

What is claimed is: 
     
         1 . A deimmunized variant albumin-binding domain (ABD) of a wild-type ABD of a Streptococcal Protein G, wherein the amino acid sequence of said deimmunized variant ABD differs from the amino acid sequence of SEQ ID NO:304 in comprising:
 (A) a leucine to alanine substitution at position 24 of SEQ ID NO:304, an isoleucine to alanine substitution at position 25 of SEQ ID NO:304, and a valine to alanine substitution at position 31 of SEQ ID NO:304; or   (B) an asparagine to aspartic acid substitution at position 26 of SEQ ID NO:304, a threonine to serine substitution at position 30 of SEQ ID NO:304, and a valine to alanine substitution at position 31 of SEQ ID NO:304.   
     
     
         2 . The deimmunized variant ABD of  claim 1 , wherein said deimmunized variant ABD comprises said leucine to alanine substitution at position 24 of SEQ ID NO:304, said isoleucine to alanine substitution at position 25 of SEQ ID NO:304, and said valine to alanine substitution at position 31 of SEQ ID NO:304. 
     
     
         3 . The deimmunized variant ABD of  claim 1 , wherein said deimmunized variant ABD comprises said asparagine to aspartic acid substitution at position 26 of SEQ ID NO:304, said threonine to serine substitution at position 30 of SEQ ID NO:304, and said valine to alanine substitution at position 31 of SEQ ID NO:304. 
     
     
         4 . The deimmunized variant ABD of  claim 1 , wherein said deimmunized variant ABD has the sequence of SEQ ID NO:328 or SEQ ID NO:329. 
     
     
         5 . A fusion protein comprising the deimmunized variant ABD of  claim 1 , covalently linked to a heterologous polypeptide, wherein the serum half-life of said fusion protein is extended relative to the serum half-life of said heterologous polypeptide not covalently linked to said deimmunized variant ABD. 
     
     
         6 . A fusion protein comprising the deimmunized variant ABD of  claim 4 , covalently linked to a heterologous polypeptide, wherein the serum half-life of said fusion protein is extended relative to the serum half-life of said heterologous polypeptide not covalently linked to said deimmunized variant ABD. 
     
     
         7 . The fusion protein of  claim 5 , further comprising a second deimmunized variant ABD of  claim 1  covalently linked to said heterologous polypeptide, wherein said second deimmunized variant ABD is the same or a different deimmunized variant ABD. 
     
     
         8 . The deimmunized variant ABD of  claim 1 , wherein said deimmunized variant ABD is covalently linked to an antigen-binding molecule, and wherein the serum half-life of antigen-binding molecule is extended relative to the serum half-life of said antigen-binding molecule not covalently linked to said deimmunized variant ABD. 
     
     
         9 . The deimmunized variant ABD of  claim 4 , wherein said deimmunized variant ABD is covalently linked to an antigen-binding molecule, and wherein the serum half-life of antigen-binding molecule is extended relative to the serum half-life of said antigen-binding molecule not covalently linked to said deimmunized variant ABD. 
     
     
         10 . The deimmunized variant ABD of  claim 8 , wherein said antigen-binding molecule is an antibody, an antigen-binding fragment thereof, an scFv, or a diabody. 
     
     
         11 . The deimmunized variant ABD of  claim 9 , wherein said antigen-binding molecule is an antibody, an antigen-binding fragment thereof, an scFv, or a diabody. 
     
     
         12 . The deimmunized variant ABD of  claim 8 , wherein said antigen is a breast cancer antigen, an ovarian cancer antigen, a prostate cancer antigen, a cervical cancer antigen, a pancreatic carcinoma antigen, a lung cancer antigen, a bladder cancer antigen, a colon cancer antigen, a testicular cancer antigen, a glioblastoma cancer antigen, an antigen associated with a B cell malignancy, an antigen associated with multiple myeloma, an antigen associated with non-Hodgkin's lymphoma, or an antigen associated with chronic lymphocytic leukemia. 
     
     
         13 . The deimmunized variant ABD of  claim 9 , wherein said antigen is a breast cancer antigen, an ovarian cancer antigen, a prostate cancer antigen, a cervical cancer antigen, a pancreatic carcinoma antigen, a lung cancer antigen, a bladder cancer antigen, a colon cancer antigen, a testicular cancer antigen, a glioblastoma cancer antigen, an antigen associated with a B cell malignancy, an antigen associated with multiple myeloma, an antigen associated with non-Hodgkin's lymphoma, or an antigen associated with chronic lymphocytic leukemia. 
     
     
         14 . A method for extending the serum half-life of a polypeptide, which comprises covalently linking said polypeptide to a deimmunized variant albumin-binding domain (ABD) of a wild-type ABD of a Streptococcal Protein G, wherein the amino acid sequence of said deimmunized variant ABD differs from the amino acid sequence of SEQ ID NO:304 in comprising:
 (A) a leucine to alanine substitution at position 24 of SEQ ID NO:304; an isoleucine to alanine substitution at, position 25 of SEQ ID NO:304; and a valine to alanine substitution at position 31 of SEQ ID NO:304; or   (B) an asparagine to aspartic acid substitution at position 26 of SEQ ID NO:304; a threonine to serine substitution at position 30 of SEQ ID NO:304; and a valine to alanine substitution at position 31 of SEQ ID NO:304,   said extension of serum half-life being relative to the serum half-life of said polypeptide if lacking said deimmunized variant ABD.   
     
     
         15 . The method of  claim 14 , wherein said deimmunized variant ABD comprises said leucine to alanine substitution at position 24 of SEQ ID NO:304; said isoleucine to alanine substitution at, position 25 of SEQ ID NO:304; and said valine to alanine substitution at position 31 of SEQ ID NO:304. 
     
     
         16 . The method of  claim 14 , wherein said deimmunized variant ABD comprises said asparagine to aspartic acid substitution at position 26 of SEQ ID NO:304;
 said threonine to serine substitution at position 30 of SEQ ID NO:304; and said valine to alanine substitution at position 31 of SEQ ID NO:304.   
     
     
         17 . The method of  claim 14 , wherein said deimmunized variant ABD has the sequence of SEQ ID NO:328 or SEQ ID NO:329. 
     
     
         18 . A composition comprising the fusion protein of  claim 5  and a pharmaceutically acceptable carrier. 
     
     
         19 . A composition comprising the fusion protein of  claim 6  and a pharmaceutically acceptable carrier. 
     
     
         20 . A composition comprising the fusion protein of  claim 7  and a pharmaceutically acceptable carrier. 
     
     
         21 . A composition comprising the deimmunized variant ABD of  claim 8  and a pharmaceutically acceptable carrier. 
     
     
         22 . A composition comprising the deimmunized variant ABD of  claim 9  and a pharmaceutically acceptable carrier. 
     
     
         23 . An expression vector encoding the deimmunized variant ABD of  claim 1 . 
     
     
         24 . An expression vector encoding the deimmunized variant ABD of  claim 4 . 
     
     
         23 . An expression vector encoding the fusion protein of  claim 5 . 
     
     
         24 . An expression vector encoding the fusion protein of  claim 6 .