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B‐cell epitopes on the hepatitis C virus nucleocapsid protein determined by human monospecific antibodies
Author(s) -
Akatsuka Toshitaka,
Donets Mickhail,
Scaglione Luca,
Ching WeiMei,
WaiKuo Shih J.,
Di Bisceglie Adrian M.,
Feinstone Stephen M.
Publication year - 1993
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840180306
Subject(s) - epitope , virology , antibody , recombinant dna , hepatitis c virus , microbiology and biotechnology , virus , biology , peptide sequence , amino acid , hepatitis b virus , peptide , biochemistry , gene , immunology
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. (HEPATOLOGY 1993;18:503–510.)