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