Albumin variants

Abstract

The present invention relates to variants of a parent albumin, the variants having altered plasma half-life compared with the parent albumin. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

1 . (canceled) 
     
     
         2 . A conjugate comprising:
 a polypeptide comprising an albumin having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 2 and having a substitution in Domain I of said albumin and having a substitution in Domain III of said albumin relative to the amino acid sequence set forth in SEQ ID NO: 2, wherein said albumin has an increased binding affinity to FcRn relative to the binding affinity of an albumin comprising the amino acid sequence of SEQ ID NO: 2 to FcRn, and wherein said substitution in Domain I is selected from amino acids corresponding to positions 82, 83, 111, 112, or any combination thereof of the amino acid sequence of SEQ ID NO: 2 and said substitution in Domain III is selected from amino acids corresponding to positions 425, 505, 510, 512, 524, 527, 531, 534, 569, 573, 575, or any combination thereof of the amino acid sequence of SEQ ID NO: 2; and   a conjugation partner.   
     
     
         3 . The conjugate of  claim 2 , wherein the conjugation partner is a therapeutic, prophylactic, diagnostic, imaging, or other moiety. 
     
     
         4 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions (a) 111 and 573; (b) 82 and 83; (c) 82 and 111; (d) 82 and 112; (e) 82 and 573; (f) 83 and 111; (g) 83 and 112; (h) 83 and 573; (i) 111 and 112; (j) 83, 111, and 573; (k) 112 and 573; (l) 82, 83, and 111; (m) 82, 83, and 112; (n) 82, 83, and 573; (o) 82, 111, and 112; (p) 82, 111, and 573; (q) 82, 112, and 573; (r) 83, 111, and 112; (s) 83, 112, and 573; (t) 111, 112, and 573; (u) 82, 83, 111, and 112; (v) 82, 83, 111, and 573; (w) 82, 83, 112, and 573; (x) 82, 111, 112, and 573; (y) 83, 111, 112, and 573; or (z) 82, 83, 111, 112, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         5 . The conjugate of  claim 2 , wherein said substitution in Domain I is selected from amino acids corresponding to positions 83 or 111 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         6 . The conjugate of  claim 2 , wherein said substitution in Domain III is at the amino acid corresponding to position 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         7 . The conjugate of  claim 2 , wherein said substitution in Domain I is at the amino acid corresponding to position 83 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is an asparagine, lysine, or serine. 
     
     
         8 . The conjugate of  claim 2 , wherein said substitution in Domain I is at the amino acid corresponding to position 111 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is an aspartic acid, glycine, histidine, arginine, glutamine, or glutamic acid. 
     
     
         9 . The conjugate of  claim 2 , wherein said substitution in Domain III is at the amino acid corresponding to position 573 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is a proline, tyrosine, tryptophan, histidine, phenylalanine, threonine, isoleucine, or valine. 
     
     
         10 . The conjugate of  claim 2 , wherein said substitution in Domain III is at the amino acid corresponding to position 573 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is a proline, tyrosine, or tryptophan. 
     
     
         11 . The conjugate of  claim 2 , wherein said substitution in Domain III is the amino acid corresponding to at position 573 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is a proline. 
     
     
         12 . The conjugate of  claim 2 , wherein the polypeptide has an increased plasma half-life compared with the half-life of the albumin comprising the amino acid sequence of SEQ ID NO: 2. 
     
     
         13 . The conjugate of  claim 2 , wherein said polypeptide has a stronger binding affinity to FcRn and optionally, a longer plasma half-life relative to a polypeptide comprising the amino acid sequence of SEQ ID NO: 2. 
     
     
         14 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 111 and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         15 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 83, 111, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         16 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 82, 111, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         17 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 111, 112, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         18 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 82, 83, 111, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         19 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 82, 111, 112, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         20 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 83, 111, 112, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         21 . The conjugate of  claim 2 , wherein the polypeptide comprises substitutions at amino acids corresponding to positions 82, 83, 111, 112, and 573 of the amino acid sequence of SEQ ID NO: 2. 
     
     
         22 . The conjugate of  claim 2 , wherein said substitution in Domain I is at the amino acid corresponding to position 83 of the amino acid sequence SEQ ID NO: 2 and wherein said substitution is an asparagine. 
     
     
         23 . The conjugate of  claim 2 , wherein said substitution in Domain I is at the amino acid corresponding to position 83 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is a lysine. 
     
     
         24 . The conjugate of  claim 2 , wherein the said substitution in Domain I is at the amino acid corresponding to position 111 of the amino acid sequence of SEQ ID NO: 2 and, wherein said substitution is a glutamic acid. 
     
     
         25 . The conjugate of  claim 2 , wherein said substitution in Domain I is at the amino acid corresponding to positions 83 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is a lysine or asparagine, and the said substitution in Domain III is at the amino acid corresponding to position 573 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is a proline. 
     
     
         26 . The conjugate of  claim 2 , wherein said substitutions in Domain I are at the amino acids corresponding to positions 83 and 111 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitutions are an asparagine and a glutamic acid, respectively, and the said substitution in Domain III is at the amino acid corresponding to position 573 of the amino acid sequence of SEQ ID NO: 2 and wherein said substitution is a proline. 
     
     
         27 . A method of producing a conjugate comprising:
 providing an albumin having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 2 and having a substitution in Domain I of said albumin and having a substitution in Domain III of said albumin relative to the amino acid sequence set forth in SEQ ID NO: 2, wherein said albumin has an increased binding affinity to FcRn relative to the binding affinity of an albumin comprising the amino acid sequence of SEQ ID NO: 2 to FcRn, and wherein said substitution in Domain I is selected from amino acids corresponding to positions 82, 83, 111, 112, or any combination thereof of the amino acid sequence of SEQ ID NO: 2 and said substitution in Domain III is selected from amino acids corresponding to positions 425, 505, 510, 512, 524, 527, 531, 534, 569, 573, 575, or any combination thereof of the amino acid sequence of SEQ ID NO: 2; and   conjugating the albumin to a conjugation partner selected from a therapeutic, prophylactic, diagnostic, imaging or other moiety.