ISOLATION AND CHARACTERIZATION OF HUMAN MONOCLONAL-ANTIBODIES AGAINSTHEPATITIS-C VIRUS ENVELOPE GLYCOPROTEINS

The isolation and characterization of human monoclonal antibodies (hum Abs) against the hepatitis C Virus (HCV) glycoproteins El and E2 are d escribed. B-cells from blood donors with anti-HCV were transformed wit h 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 E l 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 humA bs was determined both by ELISA and Western blot assays. Several recom binant extracts expressing either the El or E2 protein or truncated fo rms were used in an attempt to map the epitopes on the viral glycoprot eins. Some of the humAbs were used successfully for immunofluorescence investigation of transfected cells. Seven specific anti-E2 humAbs, wh ich react with the envelope protein 2 of genotype 1a and 1b isolates, were characterized. (C) 1998 Wiley-Liss, Inc.