Synthetic antigens for the detection of antibodies to hepatitis C virus

Abstract

Peptide sequences are provided which are capable of mimicking proteins encoded by HCV for use as reagents for screening of blood and blood products for prior exposure to HCV. The peptides are at least 5 amino acids long and can be used in various specific assays for the detection of antibodies to HCV, for the detection of HCV antigens, or as immunogens.

1 - 25 . (Canceled)  
     
     
         26 . A kit for the detection of anti-hepatitis C virus antibodies in a body fluid sample, comprising: 
 at least one epitope selected from the group consisting of:    (a) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1688 to 1707 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (b) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1694 to 1713 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (c) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1706 to 1725 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (d) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1712 to 1731 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (e) at least 5 to at most 12 amino acids located in the region consisting of amino acids 1718 to 1737 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (f) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1724 to 1743 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (g) at least 5 to at most 12 amino acids located in the region consisting of amino acids 1730 to 1749 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (h) at least 5 to at most 20 amino acids located in the region consisting of amino acids 2287 to 2306 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (i) at least 5 to at most 20 amino acids located in the region consisting of amino acids 2299 to 2318 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (j) at least 5 to at most 20 amino acids located in the region consisting of amino acids 2311 to 2330 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (k) at least 5 to at most 20 amino acids located in the region consisting of amino acids 7 to 26 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (m) at least 5 to at most 20 amino acids located in the region consisting of amino acids 13 to 32 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV, and    (n) at least 5 to at most 20 amino acids located in the region consisting of amino acids 49 to 68 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV; and    a means for detecting an immunological complex formed between said epitope and said antibodies.    
     
     
         27 . The kit of  claim 26 , comprising a collection of at least two epitopes selected from the group consisting of (a), (b), (c), (d), (e) and (g).  
     
     
         28 . The kit of  claim 27 , comprising a collection of epitopes (a), (b), (c), (d), (e) and (g).  
     
     
         29 . The kit of  claim 26 , comprising a collection of at least two epitopes selected from the group consisting of (a), (b), (c), (d), (e) and (g) and at least two epitopes selected from the group consisting of (k), (m) and (n).  
     
     
         30 . The kit of  claim 29 , comprising a collection of epitopes selected from the group consisting of (a), (b), (c), (d), (e), (g), (k), (m) and (n).  
     
     
         31 . The kit of  claim 26 , comprising a collection of at least two epitopes selected from the group consisting of (a), (b), (c), (d), (e) and (g) and at least two epitopes selected from the group consisting of (k), (m) and (n), and at least two epitopes selected from the group consisting of (h), (i) and (j).  
     
     
         32 . The kit of  claim 31 , comprising a collection of epitopes selected from the group consisting of (a), (b), (c), (d), (e), (g), (k), (m), (n), (h), (i) and (j).  
     
     
         33 . The kit of  claim 26 , comprising a collection of at least two epitopes selected from the group consisting of (b), (c) and (d), and an epitope (m), and at least two epitopes selected from the group consisting of (h), (i) and (j).  
     
     
         34 . The kit of  claim 33 , comprising a collection of epitopes selected from the group consisting of (b), (c), (d), (m), (h), (i) and (j).  
     
     
         35 . The kit according to any one of claims  26 - 34  wherein said epitopes are independently produced by recombinant expression or chemical synthesis.  
     
     
         36 . The kit according to any one of claims  27 - 32  wherein said epitopes are produced by recombinant expression or chemical synthesis.  
     
     
         37 . The kit according to any one of claims  33 - 34  wherein at least one of said epitopes are produced by chemical synthesis.  
     
     
         38 . The kit of  claim 26 , comprising a collection of epitopes selected from the group consisting of (k), (m), (n), (b), and (i).  
     
     
         39 . The kit of  claim 26 , comprising a collection of epitopes selected from the group consisting of (k), (n), (b), (d) and (i).  
     
     
         40 . The kit of  claim 26 , comprising a collection of epitopes selected from the group consisting of (k), (m), (n), (a), (d) and (i).  
     
     
         41 . The kit of  claim 26 , comprising a collection of epitopes selected from the group consisting of (k), (b) and (i).  
     
     
         42 . The kit of  claim 26 , comprising a collection of epitopes selected from the group consisting of (k), (m) and (n).  
     
     
         43 . The kit of  claim 26 , comprising a collection of epitopes selected from the group consisting of (a), (b), (d) and (i).  
     
     
         44 . The kit of  claim 26 , comprising a collection of epitopes selected from the group consisting of (h), (i) and (j).  
     
     
         45 . The kit of  claim 26 , wherein said epitope is selected from the group consisting of (b), (i) and (m).  
     
     
         46 . The kit according to any one of claims  38 - 45  wherein said epitopes are independently produced by recombinant expression or chemical synthesis.  
     
     
         47 . A device for detecting anti-hepatitis C virus antibodies, comprising: 
 (i) an isolated body fluid sample comprising said anti-hepatitis C virus antibodies,    (ii) at least one epitope selected from the group consisting of:    (a) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1688 to 1707 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (b) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1694 to 1713 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (c) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1706 to 1725 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (d) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1712 to 1731 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (e) at least 5 to at most 12 amino acids located in the region consisting of amino acids 1718 to 1737 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (f) at least 5 to at most 20 amino acids located in the region consisting of amino acids 1724 to 1743 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (g) at least 5 to at most 12 amino acids located in the region consisting of amino acids 1730 to 1749 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (h) at least 5 to at most 20 amino acids located in the region consisting of amino acids 2287 to 2306 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (i) at least 5 to at most 20 amino acids located in the region consisting of amino acids 2299 to 2318 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (j) at least 5 to at most 20 amino acids located in the region consisting of amino acids 2311 to 2330 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (k) at least 5 to at most 20 amino acids located in the region consisting of amino acids 7 to 26 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV,    (m) at least 5 to at most 20 amino acids located in the region consisting of amino acids 13 to 32 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV, and    (n) at least 5 to at most 20 amino acids located in the region consisting of amino acids 49 to 68 of the HCV polyprotein of an HCV isolate wherein said epitope is capable of providing for immunological competition with at least one strain of HCV;    wherein said at least one epitope is bound to an anti-hepatitis C virus antibody contained in said body sample in the form of an immunological complex.    
     
     
         48 . An immunological complex comprising an epitope of any of claims  27  and  45  or a collection of epitopes of any of claims  28 - 34  and  38 - 44 , and a an anti-hepatitis C virus antibody contained in an isolated body fluid sample.  
     
     
         49 . A device for detecting anti-hepatitis C virus antibodies, comprising: 
 (i) an isolated body fluid sample comprising said anti-hepatitis C virus antibodies, and    (ii) at least one epitope collection of any of claims  28 - 34  and  38 - 44 , wherein at least one epitope of said at least one epitope collection is bound to an anti-hepatitis C virus antibody contained in said body sample in the form of an immunological complex.    
     
     
         50 . A method for the detection of antibodies to hepatitis C virus present in a body fluid sample comprising the steps of: 
 (a) contacting a body fluid sample of a person to be diagnosed with at least one epitope of any of claims  27  and  45  or a collection of epitopes of any of claims  28 - 34  and  38 - 44 , and    (b) detecting an immunological complex formed between antibodies in said body fluid sample and said at least one epitope or an epitope of said collection of epitopes, as an indication of the presence of antibodies to hepatitis C virus in said body fluid sample.    
     
     
         51 . The kit of any one of claims  26 - 34  and  38 - 44 , wherein said at least one epitope or combination of epitopes are bound to a nylon membrane or microtiter plate.  
     
     
         52 . The kit of  claim 51  wherein said at least one epitope or combination of epitopes are indirectly bound to said nylon membrane or microtiter plate.  
     
     
         53 . The immunological complex of  claim 48  bound to a nylon membrane or microtiter plate.  
     
     
         54 . A combination of hepatitis C viral (HCV) epitopes comprising 
 (a) a first HCV epitope comprising at least 5 amino acids from at least one first domain selected from the group consisting of the amino acids 1-20, 7-26, 8-18,13-32, 37-56, 49-68, 61-80 and 73-92 of the HCV polyprotein; and    (b) a second HCV epitope comprising at least 5 amino acids from at least one second domain selected from the group consisting of amino acids 1688-1707, 1694-1713, 1706-1725, 1712-1731, 1718-1737, 1724-1743, 1730-1749, 2263-2282, 2275-2294, 2287-2306, 2299-2318, and 2311-2330 of the HCV polyprotein.    
     
     
         55 . A combination of hepatitis C viral (HCV) epitopes comprising 
 (a) a first HCV epitope comprising at least 5 amino acids from at least one first domain selected from the group consisting of the amino acids 1-20, 7-26, 8-18, 13-32, 37-56, 49-68, 61-80 and 73-92 of the HCV polyprotein; and    (b) a second HCV epitope comprising at least 5 amino acids from at least one second domain selected from the group consisting of amino acids 1688-1707, 1694-1713, 1706-1725, 1712-1731, 1718-1737, 1724-1743 and 1730-1749 of the HCV polyprotein, and    (c) a third HCV epitope comprising at least 5 amino acids from at least one third domain selected from the group consisting of amino acids 2263-2282, 2275-2294, 2287-2306, 2299-2318, and 2311-2330 of the HCV polyprotein.    
     
     
         56 . A combination of  claim 54  or  55  wherein said first domain is selected from the group consisting of amino acids 7-26, 13-32, and 37-56, 49-68 of the HCV polyprotein.  
     
     
         57 . A combination of  claim 54  wherein said second domain is selected from the group consisting of amino acids 1688-1707, 1694-1713, 1706-1725, 1712-1731, 1718-1737, 1724-1743, 1730-1749, 2287-2306, 2299-2318, and 2311-2330 of the HCV polyprotein  
     
     
         58 . A combination of  claim 55  wherein said second domain is selected from the group consisting of amino acids 1688-1707, 1694-1713, 1706-1725, 1712-1731, 1718-1737, 1724-1743, and 1730-1749, of the HCV polyprotein and said third domain is selected from the group consisting of amino acids 2287-2306, 2299-2318, and 2311-2330 of the HCV polyprotein.  
     
     
         59 . A combination according to  claim 54 ,  55 ,  57 , or  58 , wherein said HCV epitopes are individually produced by recombinant expression or chemical synthesis.  
     
     
         60 . A combination according to  claim 54 ,  55 ,  57 , or  58 , wherein the combination is in the form of a fusion polypeptide.  
     
     
         61 . A combination according to  claim 54 ,  55 ,  57 , or  58 , wherein said epitopes are bound to a solid surface.  
     
     
         62 . A combination according to claims  54 ,  55 ,  57 , or  58 , wherein the combination is packaged into a kit further comprising control reagents for detecting antibodies to hepatitis C virus (HCV) in a mammalian body component suspected of containing said antibodies.  
     
     
         63 . A method of designing a kit for detection of anti-hepatitis C virus antibodies in a body fluid sample, comprising selecting at least one epitope of  claim 26  and combining said at least one epitope with a support suitable for detecting said antibodies bound to said epitope.  
     
     
         64 . A combination according to  claim 56 , wherein said HCV epitopes are individually produced by recombinant expression or chemical synthesis.  
     
     
         65 . A combination according to  claim 56 , wherein the combination is in the form of a fusion polypeptide.  
     
     
         66 . A combination according to  claim 56 , wherein said epitopes are bound to a solid surface.  
     
     
         67 . A combination according to  claim 56 , wherein the combination is packaged into a kit further comprising control reagents for detecting antibodies to hepatitis C virus (HCV) in a mammalian body component suspected of containing said antibodies.