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 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.