Serum albumin-binding immunoglobulin variable domains

Abstract

The present invention relates to amino acid sequences binding to serum albumin. In particular, the present invention relates to improved immunoglobulin single variable domains (also referred to herein as “ISV's” or “ISVD's”), and more in particular improved heavy-chain immunoglobulin single variable domains, binding to serum albumin, as well as to proteins, polypeptides and other constructs, compounds, molecules or chemical entities that comprise such improved serum albumin binders.

1 .- 13 . (canceled) 
     
     
         14 . A nucleotide sequence or nucleic acid that encodes an immunoglobulin single variable domain that binds to serum albumin, wherein said immunoglobulin single variable domain comprises:
 a CDR1 according to Abm that is the following sequence: GFTFDTSSML (SEQ ID NO: 13); and   a CDR2 according to Abm that is the following sequence: VIHQSGTPTY (SEQ ID NO: 14); and   a CDR3 according to Abm that is the following sequence: FPSSRMKFDY (SEQ ID NO: 15);   
       and having:
 an amino acid residue L or V at position 11; and 
 an amino acid residue T, V or L at position 89; and 
 an amino acid residue T, K or Q at position 110; and 
 an amino acid residue S, K or Q at position 112; 
 
       such that (i) position 89 is T; or (ii) position 89 is L and position 11 is V; or (iii) position 89 is L and position 110 is K or Q; or (iv) position 89 is L and position 112 is K or Q; or (v) position 89 is L and position 11 is V and position 110 is K or Q; or (vi) position 89 is L and position 11 is V and position 112 is K or Q; or (vii) position 11 is V and position 110 is K or Q; or (viii) position 11 is V and position 112 is K or Q; wherein the positions are according to Kabat numbering. 
     
     
         15 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein position 11 is V and:
 position 89 is L; or   position 110 is K or Q; or   position 112 is K or Q; or   position 89 is L and position 110 is K or Q; or   position 89 is L and position 112 is K or Q;   
       wherein the positions are according to Kabat numbering. 
     
     
         16 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein position 89 is L and:
 position 11 is V; or   position 110 is K or Q; or   position 112 is K or Q; or   position 11 is V and position 110 is K or Q; or   position 11 is V and position 112 is K or Q;   
       wherein the positions are according to Kabat numbering. 
     
     
         17 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein position 110 is K or Q and:
 position 11 is V; or   position 89 is L; or   position 11 is V and position 89 is L;   
       wherein the positions are according to Kabat numbering. 
     
     
         18 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein position 112 is K or Q and:
 position 11 is V; or   position 89 is L; or   position 11 is V and position 89 is L;   
       wherein the positions are according to Kabat numbering. 
     
     
         19 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein position 89 is T, wherein the position is according to Kabat numbering. 
     
     
         20 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein said immunoglobulin single variable domain comprises an amino acid sequence that is chosen from the amino acid sequences of SEQ ID NOs: 72 to 99. 
     
     
         21 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein said immunoglobulin single variable domain consists of an amino acid sequence that is chosen from the following amino acid sequences: SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, or SEQ ID NO: 99. 
     
     
         22 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein said immunoglobulin single variable domain consists of an amino acid sequence that is one of the following sequences: SEQ ID NO: 78 or SEQ ID NO: 92. 
     
     
         23 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein said immunoglobulin single variable domain further comprises a C-terminal extension (X) n , in which n is 1 to 10; and each X is an amino acid residue that is independently chosen. 
     
     
         24 . The nucleotide sequence or nucleic acid according to  claim 23 , wherein each X is independently chosen from the group consisting of alanine (A), glycine (G), valine (V), leucine (L) or isoleucine (I). 
     
     
         25 . The nucleotide sequence or nucleic acid according to  claim 14 , wherein said immunoglobulin single variable domain further comprises a D at position 1 and/or an E1D mutation, wherein the position is according to Kabat numbering. 
     
     
         26 . A host or host cell containing and capable of expressing a nucleotide sequence or nucleic acid according to  claim 14 . 
     
     
         27 . A method for preparing an immunoglobulin single variable domain comprising cultivating or maintaining the host or host cell according to  claim 26  under conditions such that the host or host cell produces or expresses the immunoglobulin single variable domain. 
     
     
         28 . The method of  claim 27 , further comprising isolating the immunoglobulin single variable domain. 
     
     
         29 . A method for prevention and/or treatment of a disease or disorder using an immunoglobulin variable domain as encoded by the nucleotide sequence or nucleic acid according to  claim 14 .