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 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 valine to alanine variation at position 31 of SEQ ID NO:304; a valine to alanine variation at position 34 of SEQ ID NO:304; or an aspartate to alanine variation at position 39 of SEQ ID NO:304;
(B) a leucine to alanine variation at position 24 of SEQ ID NO:304; an isoleucine to alanine variation at, position 25 of SEQ ID NO:304; and a valine to alanine variation at position 25 of SEQ ID NO:304; or
(C) an asparagine to aspartic acid variation 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 variation at position 31 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 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.
3. The polypeptide of claim 1 , wherein said variant ABD comprises said valine to alanine variation at position 31 of SEQ ID NO:304; said valine to alanine variation at position 34 of SEQ ID NO:304; or said aspartate to alanine variation at position 39 of SEQ ID NO:304.
4. The polypeptide of claim 1 , wherein said variant ABD comprises said leucine to alanine variation at position 24 of SEQ ID NO:304; said isoleucine to alanine variation at position 25 of SEQ ID NO:304; and said valine to alanine variation at position 31 of SEQ ID NO:304.
5. The polypeptide of claim 1 , wherein said variant ABD comprises said asparagine to aspartic acid variation 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 variation at position 31 of SEQ ID NO:304.
6. The polypeptide of claim 1 , wherein said variant ABD has the sequence of SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO: 326, SEQ ID NO: 327, SEQ ID NO: 328, or SEQ ID NO: 329.
7. The polypeptide of claim 1 , wherein said polypeptide comprises an antigen-binding molecule.
8. The polypeptide of claim 7 , 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.
9. The polypeptide of claim 8 , 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.
10. The polypeptide of claim 8 , wherein said first and said second polypeptide chains are covalently linked to one another.
11. The polypeptide of claim 8 , 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.
12. The polypeptide of claim 7 , 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. 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 valine to alanine variation at position 31 of SEQ ID NO:304; a valine to alanine variation at position 34 of SEQ ID NO:304; or an aspartate to alanine variation at position 39 of SEQ ID NO:304;
(B) a leucine to alanine variation at position 24 of SEQ ID NO:304; an isoleucine to alanine variation at, position 25 of SEQ ID NO:304; and a valine to alanine variation at position 31 of SEQ ID NO:304; or
(C) an asparagine to aspartic acid variation 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 variation at position 31 of SEQ ID NO:304.
14. The method of claim 13 , 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.
15. The method of claim 13 , wherein said variant ABD comprises said valine to alanine variation at position 31 of SEQ ID NO:304; said valine to alanine variation at position 34 of SEQ ID NO:304; or said aspartate to alanine variation at position 39 of SEQ ID NO:304.
16. The method of claim 13 , wherein said variant ABD comprises said leucine to alanine variation at position 24 of SEQ ID NO:304; said isoleucine to alanine variation at position 25 of SEQ ID NO:304; and said valine to alanine variation at position 31 of SEQ ID NO:304.
17. The method of claim 16 , wherein said variant ABD comprises said asparagine to aspartic acid variation 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 variation at position 31 of SEQ ID NO:304.
18. The method of claim 13 , wherein said variant ABD has the sequence of SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO: 326, SEQ ID NO: 327, SEQ ID NO: 328, or SEQ ID NO: 329.
19. The method of claim 13 , wherein said polypeptide comprises an antigen-binding molecule.
20. The method of claim 19 , 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.
21. The method of claim 20 , 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 20 , wherein said first and said second polypeptide chains are covalently linked to one another.
23. The method of claim 20 , 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 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.
25. The polypeptide of claim 8 , 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.
26. The method of claim 20 , 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.