title: In vitro neutralization of hepatitis B virus by monoclonal antibodies against the viral surface antigen.
abstract: In vitro HBV infection and neutralization were assayed using an anti-preS1 murine monoclonal antibody (1B3) and anti-preS2 (H69K) and anti-S (CS131A) murine-human chimeric antibodies. The 1B3 (IgG1) and H69K (IgG1) was constructed previously and the CS131A was constructed for this study by expressing stably the chimeric heavy and light chains in Chinese hamster ovary cells and purifying from the culture supernatant. Previous study showed that the H69K and CS131A recognize known virus-neutralizing epitopes, while the 1B3 does not. For the assays, adult human hepatocyte primary culture was infected with the adr or ayw subtype of HBV, and the infectivity and subsequent replication was confirmed both by measuring the kinetics of HB-sAg secretion by the infected cells and detecting the intermediate replicative form of HBV DNA in the cells. Next, the hepatocytes were infected with the adr or ayw subtype of the virus that had been preincubated with various concentrations of each of the antibodies and the neutralization of HBV was analyzed. The results showed that the anti-preS2 and anti-S chimeric antibodies exhibited neutralizing activity against both the adr and ayw subtypes of the virus, with approximately 1,000 and 2,000 times higher specific activity than polyclonal hepatitis B immune globulin, respectively, but the anti-preS1 antibody scarcely neutralized the infection. The neutralizing activities of the antibodies were consistent with their epitope specificity and antigenbinding affinity, suggesting that this neutralization assay is specific. The in vitro neutralization assay will be useful for evaluating the neutralizing activity of anti-HBV antibodies before in vivo testing in chimpanzees.
authors.affiliation: Antibody Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology, Yuseong, Taejon, Korea.
authors.forename: C J
authors.initials: CJ
authors.lastname: Ryu
authors.forename: P
authors.initials: P
authors.lastname: Gripon
authors.forename: H R
authors.initials: HR
authors.lastname: Park
authors.forename: S S
authors.initials: SS
authors.lastname: Park
authors.forename: Y K
authors.initials: YK
authors.lastname: Kim
authors.forename: C
authors.initials: C
authors.lastname: Guguen-Guillouzo
authors.forename: O J
authors.initials: OJ
authors.lastname: Yoo
authors.forename: H J
authors.initials: HJ
authors.lastname: Hong
mesh_terms: D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D006510:Hepatitis B Antibodies / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D008099:Liver / Q000166:cytology; D051379:Mice; D009500:Neutralization Tests; D011233:Precipitin Tests; D011498:Protein Precursors / Q000276:immunology; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; C050806:presurface protein 1, hepatitis B surface antigen; C050807:presurface protein 2, hepatitis B surface antigen