Human monoclonal antibody fab fragments directed against hcv e2 glycoprotein and endowed with in vitro neutralizing activity

Abstract

The invention refers to a human antibody, or its functional fragments, directed against the HCV E2 glycoprotein, able to have a neutralizing activity in vivo; a composition for anti-HCV therapy comprising in a therapeutically effective amount the antibody; a composition for topical use in gel, creme, ointment and ovule formulations; the use of the antibody for validating anti-HCV vaccines.

1 . An human antibody, or its functional fragments, against the HCV E2 protein characterized in having an in vivo neutralizing activity.  
     
     
         2 . Antibody according to  claim 1 , being the antibody e137 characterized by having the following sequences of variable parts of the heavy chain and the light chain:  
       
         
           
                 
                 
               
                     
                 
                   e 137 Heavy chain (HC) 
                     
                 
                   LLEQSGSEVKVPGSSLKVSCKTSGGTFSTYTFSWVRQAPGQGLEWMGGITPIIGIA 
                 
                     
                 
                   NYARNFQDRVTITADESTSTVYMEVRRLRSEDTAVYYCAKTSEVTATRGRTFFYSA 
                 
                     
                 
                   MDVWGQGT 
                 
                     
                 
                   e 137 Light chain (LC) 
                 
                   MAELTQSPSFLSASVGDRVTITCRASQGISNYLAWYQQKPGKAPKLLIYAASTLQS 
                 
                     
                 
                   GVPSRFSGSGSWTEFTLTISRLQPEDFATYYCQHLNTYPWTFGQGT 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         3 . Antibody according to  claim 1  being the antibody e301 characterized by having the following sequences of variable parts of the heavy chain and the light chain:  
       
         
           
                 
                 
               
                     
                 
                   e 301 Heavy chain (HC) 
                     
                 
                   LLEQSGSEVKKPGSSVRVSCTTSGGTLSDYGFNWLRQAPGQGPEWMGGIIPLFRRT 
                 
                     
                 
                   TYGQKFQGRLTITADESTGATYMELSSLRSDDTAVYYCAREKVSVLTGGKSLHYFE 
                 
                     
                 
                   YWGKGT 
                 
                     
                 
                   e 301 Light chain (LC) 
                 
                   MAELTQSPATLSVSPGERATLSCRASQSVSSRLAWYQQKRGQAPSLLIYDTSSRAT 
                 
                     
                 
                   GVPARFSASGSGTQFTLTISSLQSEDFALYYCQQYNDWPSTFGQGT 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         4 . (canceled)  
     
     
         5 . (canceled)  
     
     
         6 . (canceled)  
     
     
         7 . (canceled)  
     
     
         8 . (canceled)  
     
     
         9 . (canceled)  
     
     
         10 . (canceled)  
     
     
         11 . (canceled)  
     
     
         12 . A method for validating anti-HCV vaccines using the antibody according to  claim 1 .  
     
     
         13 . A method for validating anti-HCV vaccines using the antibody according to  claim 2 .  
     
     
         14 . A method for validating anti-HCV vaccines using the antibody according to  claim 3 .  
     
     
         15 . A nucleic acid coding for the antibody according to  claim 1 .  
     
     
         16 . A nucleic acid coding for the antibody according to  claim 2 .  
     
     
         17 . A nucleic acid coding for the antibody according to  claim 3 .  
     
     
         18 . A recombinant expression vector expressing the antibody of  claim 1  in prokaryote or in eukaryote cells.  
     
     
         19 . A recombinant expression vector expressing the antibody of  claim 2  in prokaryote or in eukaryote cells.  
     
     
         20 . A recombinant expression vector expressing the antibody of  claim 3  in prokaryote or in eukaryote cells.  
     
     
         21 . A recombinant vector further comprising a nucleotide sequence coding for a signal peptide, substantially contiguous with the sequence coding for the antibody of  claim 1 , able to export this antibody out of the cell environment.  
     
     
         22 . A recombinant vector further comprising a nucleotide sequence coding for a signal peptide, substantially contiguous with the sequence coding for the antibody of  claim 2 , able to export this antibody out of the cell environment.  
     
     
         23 . A recombinant vector further comprising a nucleotide sequence coding for a signal peptide, substantially contiguous with the sequence coding for the antibody of  claim 3 , able to export this antibody out of the cell environment.  
     
     
         24 . A method of using the recombinant vector according to  claim 21  in gene therapy.  
     
     
         25 . A method of using the recombinant vector according to  claim 22  in gene therapy.  
     
     
         26 . A method of using the recombinant vector according to  claim 23  in gene therapy.  
     
     
         27 . A composition for anti-HCV therapy comprising in a therapeutically effective amount at least one of the antibodies according to the claims.  
     
     
         28 . A composition according to  claim 27  for topical use in gel, creme, ointment and ovule formulations.  
     
     
         29 . A method for the determination of the presence of antibodies directed against different epitopes of the HCV E2 protein in a biological fluid comprising the steps of: 
 a) determining the presence of antibodies in said fluid able to inhibit the binding of specific human Fab directed against different epitopes of protein E2; and    b) correlating the presence of so titered antibodies with clinical characteristics of patients, such as prognosis, responsiveness to therapy, infectivity.