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 polypeptide that comprises a portion of a deimmunized albumin-binding protein capable of binding to serum albumin; wherein said deimmunized albumin-binding protein portion is a variant of a wild-type albumin-binding domain (ABD) of a Streptococcal Protein G, said wild-type ABD having the amino acid sequence of SEQ ID NO:304; wherein said variant ABD has an amino acid sequence that differs from that of SEQ ID NO:304 in comprising:
 (A) a tyrosine to alanine variation at position 21 of SEQ ID NO:304 and an isoleucine to alanine variation at position 25 of SEQ ID NO:304; or   (B) an asparagine to serine variation at position 26 of SEQ ID NO:304 and a threonine to serine variation at position 30 of SEQ ID NO:304;   wherein said deimmunized albumin-binding protein portion extends the serum half-life of said polypeptide, relative to the serum half-life of said polypeptide if lacking said portion of said deimmunized albumin-binding protein.   
     
     
         2 . The polypeptide of  claim 1 , wherein said variant ABD comprises said tyrosine to alanine variation at position 21 of SEQ ID NO:304 and said isoleucine to alanine variation at position 25 of SEQ ID NO:304, and additionally comprises a valine to alanine variation at position 31 of SEQ ID NO:304 or an aspartate to alanine variation at position 39 of SEQ ID NO:304. 
     
     
         3 . The polypeptide of  claim 1 , wherein said variant ABD comprises said tyrosine to alanine variation at position 21 of SEQ ID NO:304 and said isoleucine to alanine variation at position 25 of SEQ ID NO:304, and additionally comprises an asparagine to serine variation at position 26 of SEQ ID NO:304 or an asparagine to aspartate variation at position 26 of SEQ ID NO:304 or an asparagine to glutamate variation at position 26 of SEQ ID NO:304. 
     
     
         4 . The polypeptide of  claim 1 , wherein said variant ABD comprises said asparagine to serine variation at position 26 of SEQ ID NO:304 and said threonine to serine variation at position 30 of SEQ ID NO:304, and additionally comprises a valine to alanine variation at position 31 of SEQ ID NO:304 or an aspartate to alanine variation at position 39 of SEQ ID NO:304. 
     
     
         5 . The polypeptide of  claim 1 , wherein said polypeptide comprises an additional portion of a deimmunized albumin-binding protein, wherein said portions of said deimmunized albumin-binding protein are both capable of binding to said serum albumin. 
     
     
         6 . The polypeptide of  claim 1 , wherein said polypeptide comprises an antigen-binding molecule. 
     
     
         7 . The polypeptide of  claim 6 , wherein said antigen-binding molecule is a diabody composed of at least a first and a second polypeptide chain which interact with one another to form two antigen-binding sites, wherein at least one of said polypeptide chains comprises said portion of said deimmunized albumin-binding protein that is capable of binding to said serum albumin. 
     
     
         8 . The polypeptide of  claim 7 , wherein both said first and said second polypeptide chains comprise said portion of said deimmunized albumin-binding protein capable of binding to said serum albumin. 
     
     
         9 . The polypeptide of  claim 7 , wherein said first and said second polypeptide chains are covalently linked to one another. 
     
     
         10 . The polypeptide of  claim 7 , wherein said diabody binds to:
 (A) the Natural Killer Group 2D (NKG2D) receptor or the T-cell receptor (TCR); and   (B) a tumor-associated antigen.   
     
     
         11 . The polypeptide of  claim 6 , 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. 
     
     
         12 . The polypeptide of  claim 7 , wherein said diabody has an antigen-binding site that binds to 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 . A method for extending the serum half-life of a polypeptide, which comprises covalently linking said polypeptide to a polypeptide portion of a deimmunized albumin-binding protein, said extension of serum half-life being relative to the serum half-life of said polypeptide if lacking said albumin-binding protein; wherein said deimmunized albumin-binding protein portion is a variant of a wild-type albumin-binding domain (ABD) of a Streptococcal Protein G, said wild-type ABD having the amino acid sequence of SEQ ID NO:304; wherein said variant ABD has an amino acid sequence that differs from that of SEQ ID NO:304 in comprising:
 (A) a tyrosine to alanine variation at position 21 of SEQ ID NO:304 and an isoleucine to alanine variation at position 25 of SEQ ID NO:304; or   (B) an asparagine to serine variation at position 26 of SEQ ID NO:304 and a threonine to serine variation at position 30 of SEQ ID NO:304.   
     
     
         14 . The method of  claim 13 , wherein said variant ABD comprises said tyrosine to alanine variation at position 21 of SEQ ID NO:304 and said isoleucine to alanine variation at position 25 of SEQ ID NO:304, and additionally comprises a valine to alanine variation at position 31 of SEQ ID NO:304 or an aspartate to alanine variation at position 39 of SEQ ID NO:304. 
     
     
         15 . The method of  claim 13 , wherein said variant ABD comprises said tyrosine to alanine variation at position 21 of SEQ ID NO:304 and said isoleucine to alanine variation at position 25 of SEQ ID NO:304, and additionally comprises an asparagine to serine variation at position 26 of SEQ ID NO:304 or an asparagine to aspartate variation at position 26 of SEQ ID NO:304 or an asparagine to glutamate variation at position 26 of SEQ ID NO:304. 
     
     
         16 . The method of  claim 13 , wherein said variant ABD comprises said asparagine to serine variation at position 26 of SEQ ID NO:304 and said threonine to serine variation at position 30 of SEQ ID NO:304, and additionally comprises a valine to alanine variation at position 31 of SEQ ID NO:304 or an aspartate to alanine variation at position 39 of SEQ ID NO:304. 
     
     
         17 . The method of  claim 13 , wherein said variant ABD comprises said tyrosine to alanine variation at position 21 of SEQ ID NO:304 and said isoleucine to alanine variation at position 25 of SEQ ID NO:304, and wherein said polypeptide comprises an additional portion of a deimmunized albumin-binding protein, wherein said portions of said deimmunized albumin-binding protein are both capable of binding to said serum albumin. 
     
     
         18 . The method of  claim 13 , wherein said polypeptide comprises an antigen-binding molecule. 
     
     
         19 . The method of  claim 18 , wherein said antigen-binding molecule is a diabody composed of at least a first and a second polypeptide chain which interact with one another to form two antigen-binding sites, wherein at least one of said polypeptide chains comprises said portion of said deimmunized albumin-binding protein that is capable of binding to said serum albumin. 
     
     
         20 . The method of  claim 18 , 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. 
     
     
         21 . The method of  claim 19 , wherein both said first and said second polypeptide chains comprise said portion of said deimmunized albumin-binding protein capable of binding to said serum albumin. 
     
     
         22 . The method of  claim 19 , wherein said first and said second polypeptide chains are covalently linked to one another. 
     
     
         23 . The method of  claim 19 , wherein said diabody binds to:
 (A) the Natural Killer Group 2D (NKG2D) receptor or the T-cell receptor (TCR); and   (B) a tumor-associated antigen.   
     
     
         24 . The method of  claim 19 , wherein said diabody has an antigen binding site that binds to 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.