HCV core protein sequences
View Patent ↗The present invention describes peptides and recombinant proteins containing Hepatitis C virus core protein sequence in which one or more of the amino acids have been modified or deleted to remove the ability of these proteins to bind to specific anti-HCV murine monoclonal antibodies. The deletions and modifications are designed as to maintain the ability of this protein to be used in immunoassays used for the detection of anti-HCV antibodies in individuals infected with HCV.
1. A kit for the simultaneous detection of HCV antibody and HCV antigen comprising
A) a solid phase coated with
1) an HCV core peptide, comprising the amino acid sequence of SEQ ID NO: 19, wherein at least two of the following amino acid segments have one or more amino acids deleted or one or more amino acids altered,
i) MSTNPKPQRK (residues 1-10);
ii) IVGGVYLL (residues 30-37);
iii) RLGVRATR (residues 43-50); and
iv) AQPGYPWPLYGNEGCGWAGWLLSPRGSRPSWGPTDPRRRSRNLGKV IDTLTCGF (residues 77-130);
2) HCV non-core protein, and
3) at least one anti-HCV monoclonal antibody that binds to HCV core antigen and does not bind with the HCV core peptide described in A) 1); and
B) a labeled anti-HCV monoclonal antibody that binds to HCV core antigen and does not bind to the HCV core peptide described in A) 1) and does not bind to the same epitope as the anti-HCV monoclonal antibody described in A) 3).
2. The kit according to claim 1 wherein the HCV non-core protein is selected from c200 and NS5.
3. The kit according to claim 1 wherein the labeled anti-HCV monoclonal antibody is labeled with an enzyme, a fluorophore, or a radioactive label.
4. The kit according to claim 1 wherein the solid phase is a microtitre well, polystyrene bead, magnetic bead or membrane.
5. An isolated HCV core peptide, comprising the amino acid sequence of SEQ ID NO: 19, wherein at least two of the following segments of amino acids have one or more amino acids deleted or one or more amino acids altered,
i) MSTNPKPQRK (residues 1-10);
ii) IVGGVYLL (residues 30-37);
iii) RLGVRATR (residues 43-50); and
iv) AQPGYPWPLYGNEGCGWAGWLLSPRGSRPSWGPTDPRRRSRNLGKV IDTLTCGF (residues 77-130),
and wherein amino acids 11-29, 38-42, and 51-76 are not deleted or altered.
6. The isolated HCV core peptide claimed in claim 5 wherein amino acids MSTNPKPQ have been deleted from section i) and amino acids VGG have been deleted from section ii).