Domain antibody construct

Abstract

The present invention provides a domain antibody construct which binds to human TNF-α, with the construct comprising: (a) a domain antibody (dAb) which binds to human TNF-α; (b) a modified hinge region sequence; (c) a human or primate heavy chain constant region sequence having a truncated C H 1 domain of not more than 20 residues, wherein the modified hinge region sequence contains either a deletion or a single amino acid substitution of at least one cysteine residue which normally facilitates disulfide bond formation between heavy and light antibody chains.

1 . A domain antibody construct which binds to human TNF-α, the construct comprising:
 (a) a domain antibody (dAb) which binds to human TNF-α; 
 (b) a modified hinge region sequence; 
 (c) a human or primate heavy chain constant region sequence having a truncated C H 1 domain of not more than 20 residues, 
 wherein said modified hinge region sequence contains either a deletion or a single amino acid substitution of at least one cysteine residue which normally facilitates disulfide bond formation between heavy and light antibody chains. 
 
     
     
         2 . The domain antibody construct according to  claim 1  wherein the human or primate heavy chain constant region sequence having a truncated C H 1 domain comprises not more than 10 residues. 
     
     
         3 . The domain antibody construct according to  claim 2  wherein the human or primate heavy chain constant region sequence having a truncated C H 1 domain comprises not more than 5 residues. 
     
     
         4 . The domain antibody construct according to  claim 3  wherein the human or primate heavy chain constant region sequence having a truncated C H 1 domain comprises not more than a single residue. 
     
     
         5 . The domain antibody construct according to  claim 1  wherein the sequence of the C H 1 domain and the hinge region is XEPKSZDKTHTCPPCPA (SEQ ID No: 64) wherein X is valine, leucine or isoleucine and Z is absent or an amino acid other than cysteine. 
     
     
         6 . The domain antibody construct according to  claim 5  wherein X is valine and Z is serine. 
     
     
         7 . The domain antibody construct according to  claim 1  wherein the dAb comprises an immunoglobulin heavy or light chain variable domain, wherein said variable domain comprises at least one complementarity determining region (CDR) having a sequence derived from a New World primate wherein the CDR is selected from the group consisting of AATKLQS (SEQ ID No:1), EASSLQS (SEQ ID No:2), EASKLQS (SEQ ID No:3), and SASNLET (SEQ ID No:4). 
     
     
         8 . The domain antibody construct according to  claim 7  wherein the CDR is CDR2. 
     
     
         9 . The domain antibody construct according to  claim 1  wherein the domain antibody has a sequence selected from the group consisting of: 
       
         
           
                 
               
                   (SEQ ID No: 7) 
                 
                   DIQMTQSPSSLSASVGDRVTITCRASQSIDSYLHWYQQKPGKAPKWYS 
                 
                     
                 
                   ASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVVWRPFTF 
                 
                     
                 
                   GQGTKVEIKR; 
                 
                     
                 
                   (SEQ ID No: 8) 
                 
                   DIQMTQSPSSLSASVGDRVTITCRASQAIDSYLHWYQQKPGKAPKLLI 
                 
                     
                 
                   YSASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVVWRPF 
                 
                     
                 
                   TFGQGTKVEIKR; 
                 
                     
                 
                   (SEQ ID No: 9) 
                 
                   DIQMTQSPSSLSASVGDRVTITCRASQSIDSYLHWYQQKPGKAPKLLI 
                 
                     
                 
                   YSASNLETGVPSRFSGSGSGTDFTLTISSLLPEDFATYYCQQVVWRPF 
                 
                     
                 
                   TFGQGTKVEIKR; 
                 
                     
                 
                   (SEQ ID No: 10) 
                 
                   DIQMTQSPSSLSASVGDRVTITCRASQAIDSYLHWYQQKPGKAPKLLI 
                 
                     
                 
                   YSASNLETGVPSRFSGSGSGTDFTLTISSLLPEDFATYYCQQVVWRPF 
                 
                     
                 
                   TFGQGTKVEIKR; 
                 
                     
                 
                   (SEQ ID No: 52) 
                 
                   DIQMTQSPSSLSASVGDRVTITCRASQSIDSYLHWYQQKPGKPPKLLI 
                 
                     
                 
                   YSASNLETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQVVWRPF 
                 
                     
                 
                   TFGQGTKVEIKR; 
                 
                     
                 
                   (SEQ ID No: 53) 
                 
                   DIQMTQSPSSLSASVGDRVTITCRASQSIDSYLHWYQQKPGKAPKLLI 
                 
                     
                 
                   YSASNLETGVPSRFSGRGSGTDFTLTISSLQPEDFATYYCQQVVWRPF 
                 
                     
                 
                   TFGQGTKVEIKR; 
                 
                     
                 
                   (SEQ ID No: 54)  
                 
                   DIQMTQSPSSLSASVGDRVTITCRASQSIDSYLHWYQQKPGKAPKLLI 
                 
                     
                 
                   YSASNLETGVPSRFSGSGSGTDFTLTISSLVPEDFATYYCQQVVWRPF 
                 
                     
                 
                   TFGQGTKVEIKR; 
                 
                   and 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       a sequence at least 95% identical to one of these sequences. 
     
     
         10 . The domain antibody construct according to  claim 1  wherein the cysteine residue within the hinge region which normally facilitates disulfide bond formation between heavy and light antibody chains is substituted with a serine residue. 
     
     
         11 . The domain antibody construct according to  claim 1  wherein the hinge region comprises the sequence EPKSSDKTHTCPPCPA (SEQ ID No:12). 
     
     
         12 - 20 . (canceled) 
     
     
         21 . The domain antibody construct according to  claim 1  wherein the amino acid sequence is at least 60% identical to the sequence set forth in SEQ ID No:11. 
     
     
         22 - 29 . (canceled) 
     
     
         30 . A dimeric domain antibody construct which binds to human TNF-α wherein the dimer consists of two domain antibody constructs according to  claim 1 . 
     
     
         31 . The dimeric domain antibody construct according to  claim 30  wherein the dimeric domain antibody construct is a homodimer. 
     
     
         32 . The dimeric domain antibody construct according to  claim 31  wherein the domain antibody constructs making up the homodimer comprises an amino acid sequence which is at least 60% identical to the sequence set forth in SEQ ID No:11. 
     
     
         33 - 50 . (canceled) 
     
     
         51 . A pharmaceutical composition comprising an effective amount of a domain antibody construct according to  claim 1 , together with a pharmaceutically acceptable carrier or diluent. 
     
     
         52 . A pharmaceutical composition comprising an effective amount of a dimeric domain antibody construct according to  claim 30 , together with a pharmaceutically acceptable carrier or diluent. 
     
     
         53 - 59 . (canceled)