Premium
Isolation and characterization of human monoclonal antibodies against hepatitis C virus envelope glycoproteins
Author(s) -
da Silva Cardoso Marcia,
Siemoneit Karl,
Sturm Daniela,
Krone Christoph,
Moradpour Darius,
Kubanek Bernhard
Publication year - 1998
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/(sici)1096-9071(199805)55:1<28::aid-jmv6>3.0.co;2-q
Subject(s) - virology , monoclonal antibody , epitope , glycoprotein , biology , recombinant dna , virus , orthopoxvirus , immunofluorescence , microbiology and biotechnology , viral envelope , antibody , hepatitis c virus , western blot , vaccinia , immunology , gene , biochemistry
The isolation and characterization of human monoclonal antibodies (humAbs) against the hepatitis C Virus (HCV) glycoproteins E1 and E2 are described. B‐cells from blood donors with anti‐HCV were transformed with Epstein‐Barr virus. The supernatants of the resulting lymphoblastoid clones were screened by ELISA with an extract of cells infected with a recombinant vaccinia virus RMPA95 expressing the envelope proteins E1 and E2 of an HCV genotype 1a virus (H strain). Positive clones were fused to the heteromyeloma cell line K6H6/B5. Fifteen heterohybridoma cell lines have been established. The specificity of the isolated humAbs was determined both by ELISA and Western blot assays. Several recombinant extracts expressing either the E1 or E2 protein or truncated forms were used in an attempt to map the epitopes on the viral glycoproteins. Some of the humAbs were used successfully for immunofluorescence investigation of transfected cells. Seven specific anti‐E2 humAbs, which react with the envelope protein 2 of genotype 1a and 1b isolates, were characterized. J. Med. Virol. 55:28–34, 1998 . © 1998 Wiley‐Liss, Inc.