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B-cell epitopes on the hepatitis C virus nucleocapsid protein determined by human monospecific antibodies.

Abstract
Four monospecific antibodies against the hepatitis C virus nucleocapsid protein, which was expressed by recombinant baculovirus, were obtained by Epstein-Barr virus transformation of B cells from three patients with chronic hepatitis C virus infection. One of these antibodies was IgG and the other three were IgM. Their specificities were characterized initially by enzyme-linked immunosorbent assay and immunoblotting against hepatitis C virus proteins expressed by six recombinant baculoviruses with different hepatitis C virus sequence insertions. These specificities were confirmed, and their epitopes were more precisely determined with a series of overlapping decapeptides made by solid-phase pin technology. Two antibodies (1F4 and 2G6) reacted with the same peptides located near the amino(N)-terminus of nucleocapsid protein (amino acids 33-50). The third antibody (3B5) recognized the peptide consisting of amino acids 133-142, and the fourth antibody (3B9) was mapped to the carboxy(C)-terminus and reacted with a peptide consisting of amino acids 165-174. This epitope has not previously been reported. Two antibodies, 1F4 and 3B9, which are specific to the N-terminus and C-terminus of nucleocapsid protein, respectively, have been stably produced for more than 6 mo and are being subcloned to establish monoclonality. These antibodies should be useful reagents for the study of hepatitis C virus.
AuthorsT Akatsuka, M Donets, L Scaglione, W M Ching, J W Shih, A M Di Bisceglie, S M Feinstone
JournalHepatology (Baltimore, Md.) (Hepatology) Vol. 18 Issue 3 Pg. 503-10 (Sep 1993) ISSN: 0270-9139 [Print] United States
PMID7689529 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antibodies, Monoclonal
  • Antigen-Antibody Complex
  • Epitopes
  • Hepatitis Antibodies
  • Hepatitis C Antibodies
  • Immunoglobulin G
  • Viral Core Proteins
  • nucleocapsid protein, Hepatitis C virus
Topics
  • Antibodies, Monoclonal
  • Antigen-Antibody Complex
  • B-Lymphocytes (immunology)
  • Cell Line
  • Cell Line, Transformed
  • Epitopes (analysis)
  • Hepacivirus (immunology)
  • Hepatitis Antibodies (biosynthesis, immunology)
  • Hepatitis C (blood, immunology)
  • Hepatitis C Antibodies
  • Herpesvirus 4, Human (genetics)
  • Humans
  • Immunoglobulin G (biosynthesis, classification)
  • Viral Core Proteins (immunology)

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