US2012270208A1PendingUtilityA1

Synthetic antigens for the detection of antibodies to hepatitis c virus

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Individually held — no corporate assignee on recordPriority: Dec 14, 1990Filed: May 23, 2012Published: Oct 25, 2012
Est. expiryDec 14, 2010(expired)· nominal 20-yr term from priority
Inventors:Robert DeleysDirk PolletGeert MaertensHugo Van Heuverswjn
A61K 39/00Y10S436/82Y10S530/826C12N 2770/24222C07K 14/005
65
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Claims

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.

Claims

exact text as granted — not AI-modified
1 . A combination of distinct molecules, consisting essentially of
 (a) a first molecule which is capable of providing for immunological competition with at least one strain of HCV, comprising an amino acid sequence selected from the group consisting of at least 5 amino acids of an HCV polyprotein located in the region corresponding to amino acids 1-20 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 7-26 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 8-18 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 13-32 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 37-56 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 49-68 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 61-80 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 73-92 of SEQ ID NO:23, at least 5 amino acids of SEQ ID NO: 1, at least 5 amino acids of SEQ ID NO: 2, at least 5 amino acids of SEQ ID NO: 3, at least 5 amino acids of SEQ ID NO: 4, at least 5 amino acids of SEQ ID NO: 5, at least 5 amino acids of SEQ ID NO: 6, at least 5 amino acids of SEQ ID NO: 7, and at least 5 amino acids of SEQ ID NO:   8; and   (b) a second molecule which is capable of providing for immunological competition with at least one strain of HCV, comprising an amino acid sequence selected from the group consisting of at least 5 amino acids located in the region corresponding to amino acids 1688-1707 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 1694-1713 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 1706-1725 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 1712-1731 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 1718-1737 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 1724-1743 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 1730-1749 of SEQ ID NO:23, at least 5 amino acids of SEQ ID NO: 9, at least 5 amino acids of SEQ ID NO: 10, at least 5 amino acids of SEQ ID NO: 11, at least 5 amino acids of SEQ ID NO: 12, at least 5 amino acids of SEQ ID NO: 13, at least 5 amino acids of SEQ ID NO: 14 and at least 5 amino acids of SEQ ID NO: 15, and   (c) a third molecule which is capable of providing for immunological competition with at least one strain of HCV, comprising an amino acid sequence selected from the group consisting of at least 5 amino acids located in the region corresponding to amino acids 2263-2282 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 2275-2294 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 2287-2306 of SEQ ID NO:23, at least 5 amino acids located in the region corresponding to amino acids 2299-2318 of SEQ ID NO:23, at lease 5 amino acids located in the region corresponding to amino acids 2311-2330 of SEQ ID NO:23, at least 5 amino acids of SEQ ID NO: 16, at least 5 amino acids of SEQ ID NO: 17, at least 5 amino acids of SEQ ID NO: 18, at least 5 amino acids of SEQ ID NO: 19, and at least 5 amino acids of SEQ ID NO: 20,   wherein said molecules of (a), (b) and (c) are each separate molecules from one another, wherein said molecules of (a), (b) and (c) are each different from one another and wherein said molecules are selected from the group consisting of peptides and polypeptides, and   said combination further including at least one molecule selected from the group consisting of   SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO:   5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO:15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 7 to 26 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 13 to 32 of SEQ ID NO:23,   a peptide of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV corresponding to the region represented by amino acids 37 to 56 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 49 to 68 of SEQ ID NO:23,   a peptide of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV corresponding to the region represented by amino acids 61 to 80 of SEQ ID NO:23,   a peptide of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV corresponding to the region represented by amino acids 73 to 92 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 1688 to 1707 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 1694 to 1713 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 1706 to 1725 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids located of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV in the region corresponding to amino acids 1712 to 1731 of SEQ ID NO:23,   a peptide consisting of at least 5 to at most 12 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 1718 to 1737 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 1724 to 1743 of SEQ ID NO:23,   a peptide consisting of at least 5 to at most 12 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 1730 to 1749 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 2287 to 2306 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 2299 to 2318 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV located in the region corresponding to amino acids 2311 to 2330 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of SEQ ID NO:2,   a peptide consisting of at least 5 amino acids of SEQ ID NO:4,   a peptide consisting of at least 5, 6, 8, 12 or 20 amino acids of amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV corresponding to the region represented by 37 to 56 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of SEQ ID NO:6,   a peptide consisting of at least 5, 6, 8, 12 or 20 amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV corresponding to the region represented by amino acids 61 to 80 of SEQ ID NO:23,   a peptide consisting of at least 5, 6, 8, 12 or 20 amino acids of amino acids of the HCV polyprotein of an HCV isolate which is capable of providing for immunological competition with at least one strain of HCV corresponding to the region represented by 73 to 92 of SEQ ID NO:23,   a peptide consisting of at least 5 amino acids of SEQ ID NO:9,   a peptide consisting of at least 5 amino acids of SEQ ID NO:10,   a peptide consisting of at least 5 amino acids of SEQ ID NO:11,   a peptide consisting of at least 5 amino acids of SEQ ID NO:12,   a peptide consisting of at least 5 to at most 12 amino acids of SEQ ID NO:13,   a peptide consisting of at least 5 amino acids of SEQ ID NO:14,   a peptide consisting of at least 5 to at most 12 amino acids of SEQ ID NO:15,   a peptide consisting of at least 5 amino acids of SEQ ID NO: 18,   a peptide consisting of at least 5 amino acids of SEQ ID NO: 19, and   a peptide consisting of at least 5 amino acids of SEQ ID NO: 20.   
     
     
         2 . A combination according to  claim 1 , wherein said molecules comprising at least 5 amino acids are individually produced by recombinant expression or chemical synthesis. 
     
     
         3 . A combination according to  claim 1 , wherein one or more of said first, second, or third molecules comprises a fusion polypeptide. 
     
     
         4 . A combination according to  claim 1 , wherein the combination is packaged into a kit further comprising control reagents for detecting antibodies to hepatitis C virus (HCV) in a human body component suspected of containing said antibodies. 
     
     
         5 . A combination of  claim 1 , wherein at least one of said molecules comprises synthetic peptides or polypeptides. 
     
     
         6 . A combination of  claim 5 , wherein at least two of said molecules comprises synthetic peptides or polypeptides. 
     
     
         7 . A combination of  claim 6 , wherein at least three of said molecules comprises synthetic peptides or polypeptides. 
     
     
         8 . A combination of molecules according to  claim 1 ,
 wherein at least one of said first, second and third molecules is selected from the group consisting of:   a first molecule consisting of amino acids 1-92 of SEQ ID NO: 23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 1-92 of SEQ ID NO:23;   a second molecule consisting of amino acids 1688-1749 of SEQ ID NO: 23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 1688-1749 of SEQ ID NO: 23; and   a third molecule consisting of amino acids only 2263-2330 SEQ ID NO: 23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 2263-2330 of SEQ ID NO: 23.   
     
     
         9 . A combination of molecules according to  claim 1 , wherein at least two of said first, second and third molecules is selected from the group consisting of:
 a first molecule consisting of amino acids 1-92 of SEQ ID NO: 23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 1-92 of SEQ ID NO: 23;   a second molecule consisting of amino acids 1688-1749 of SEQ ID NO:   23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 1688-1749 of SEQ ID NO: 23; and   a third molecule consisting of amino acids 2263-2330 of SEQ ID NO: 23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 2263-2330 of SEQ ID NO: 23.   
     
     
         10 . A combination of molecules according to  claim 1 , wherein:
 said first molecule consisting of amino acids 1-92 of SEQ ID NO: 23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 1-92 of SEQ ID NO: 23;   said second molecule consisting of amino acids 1688-1749 of SEQ ID NO:   23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 1688-1749 of SEQ ID NO: 23; and   said third molecule consisting of amino acids 2263-2330 of SEQ ID NO: 23 or consisting of amino acids of an HCV polyprotein corresponding to the region represented by amino acids 2263-2330 of SEQ ID NO: 23.   
     
     
         11 . A method for making a kit for detecting human antibodies that bind to HCV amino acids, comprising combining the combination of  claim 8  with a support suitable for detecting whether antibodies bind to the molecules of said combination. 
     
     
         12 . A method for making a kit for detecting human antibodies that bind to HCV amino acids, comprising combining the combination of  claim 9  with a support suitable for detecting whether antibodies bind to the molecules of said combination. 
     
     
         13 . A method for making a kit for detecting human antibodies that bind to HCV amino acids, comprising combining the combination of  claim 10  with a support suitable for detecting whether antibodies bind to the molecules of said combination. 
     
     
         14 . A combination of  claim 1 , wherein
 said at least 5 amino acids of said first molecule are selected from the amino acids of SEQ ID NO: 2, of SEQ ID NO: 4, of SEQ ID NO: 6, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 7-26 of SEQ ID NO:23, located in the region of an HCV polyprotein corresponding to amino acids 13-32 of SEQ ID NO:23, or located in the region of an HCV polyprotein corresponding to amino acids 49-68 of SEQ ID NO:23;   said at least 5 amino acids of said second molecule are selected from the amino acids or SEQ ID NO:10, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 1694-1713 of SEQ ID NO: 23; and      wherein said at least 5 amino acids of said third molecule are selected from amino acids of SEQ ID NO:19, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 2299-2318 of SEQ ID NO:23.   
     
     
         15 . A combination of  claim 1 , wherein
 said at least 5 amino acids of said first molecule are selected from the amino acids of SEQ ID NO: 1, of SEQ ID NO: 2, of SEQ ID NO: 6, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 1-20 SEQ ID NO:23, located in the region of an HCV polyprotein corresponding to amino acids 7-26 of SEQ ID NO:23, or located in the region of an HCV polyprotein corresponding to amino acids 49-68 of SEQ ID NO:23;   said at least 5 amino acids of said second molecule are selected from the amino acids of SEQ ID NO: 10, of SEQ ID NO: 12, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 1694-1713 of SEQ ID NO:23, or located in the region of an HCV polyprotein corresponding to amino acids 1712-1731 of SEQ ID NO:23; and   wherein said at least 5 amino acids of said third molecule are selected from the amino acids of SEQ ID NO: 17, of SEQ ID NO: 19, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 2275-2294 of SEQ ID NO:23, or located in the region of an HCV polyprotein corresponding to amino acids 2299-2318 of SEQ ID NO:23.   
     
     
         16 . A combination of  claim 1 , wherein
 said at least 5 amino acids of said first molecule are selected from the amino acids of SEQ ID NO: 2, of SEQ ID NO: 4, of SEQ ID NO: 5, of SEQ ID NO: 6, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 7-26 of SEQ ID NO:23, located in the region of an HCV polyprotein corresponding to amino acids 13-32 of SEQ ID NO:23, located in the region of an HCV polyprotein corresponding to amino acids 37-56 of SEQ ID NO:23, or located in the region of an HCV polyprotein corresponding to amino acids 49-68 of SEQ ID NO:23;   said at least 5 amino acids of said second molecule are selected from the amino acids of SEQ ID NO: 9, of SEQ ID NO: 12, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 1688-1707 of SEQ ID NO:23, or located in the region of an HCV polyprotein corresponding to amino acids 1712-1731 of SEQ ID NO:23; and   wherein said at least 5 amino acids of said third molecule are selected from the amino acids of SEQ ID NO: 17, of SEQ ID NO: 19, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 2275-2294 of SEQ ID NO:23, or located in the region of amino acids 2299-2318 of SEQ ID NO:23.   
     
     
         17 . A combination of  claim 14 , wherein
 said at least 5 amino acids of said first molecule are selected from the amino acids of SEQ ID NO: 2, or from the amino acids located in the region of an HCV polyprotein corresponding to amino acids 7-26 of SEQ ID NO:23.   
     
     
         18 . A combination of  claim 1 , wherein said combination coats a well of an immunoassay plate. 
     
     
         19 . A combination of  claim 1 , wherein said combination coats a well of an immunoassay plate. 
     
     
         20 . A combination of  claim 14 , wherein said combination coats a well of an immunoassay plate. 
     
     
         21 . A combination of  claim 15 , wherein said combination coats a well of an immunoassay plate. 
     
     
         22 . A combination of  claim 16 , wherein said combination coats a well of an immunoassay plate. 
     
     
         23 . A combination of  claim 1 , wherein said combination is bound to an immunoassay strip. 
     
     
         24 . A combination of  claim 14 , wherein said combination is bound to an immunoassay strip. 
     
     
         25 . A combination of  claim 15 , wherein said combination is bound to an immunoassay strip. 
     
     
         26 . A combination of  claim 16 , wherein said combination is bound to an immunoassay strip.

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