title: Evaluation of murine monoclonal antibodies targeting different epitopes of the hepatitis B virus surface antigen by using immunological as well as molecular biology and biochemical approaches.
abstract: The hepatitis B virus surface protein (HBsAg) displays the major B cells antigenic determinants that can induce protective immunity and prevent the hepatitis B virus (HBV) infection, a major health problem. A panel of murine monoclonal antibodies against the HBsAg (MAb anti-HBs), raised after mice immunization with a pool of plasma of hepatitis chronic carriers, has been established. Mainly using simple immunological tools such as enzyme-linked immunosorbent assay (ELISA) and Western blot analysis, we could trace the location of the epitopes on the HBsAg determinants. We also report the use of two specific methodology approaches based on molecular biology and biochemical techniques such as, respectively, cloning and expression of preS1 major neutralizing epitope of the HBsAg in Escherichia coli and ELISA accomplished to chemical reduction with dithiothreitol (DTT), which were able to complete the MAb anti-HBs characterization. Our results showed that the majority of the MAbs anti-HBs were directed to the HBV common determinant a. One MAb recognizes a discontinuous epitope present in all forms of the HBsAg when evaluated by Western blot.
authors.affiliation: Laboratory of Monoclonal Antibodies Technology, Bio-Manguinhos, Oswaldo Cruz Foundation, FIOCRUZ, P.O. Box 926, Rio de Janeiro, Brazil.
authors.forename: Carlos Otávio
authors.initials: CO
authors.lastname: Alves Vianna
authors.forename: Sérgio
authors.initials: S
authors.lastname: da Silva E Mouta Júnior
authors.forename: Maria
authors.initials: M
authors.lastname: da Glória Teixeira Martins
authors.forename: Nadia Maria
authors.initials: NM
authors.lastname: Batoreu
authors.forename: João Luiz
authors.initials: JL
authors.lastname: Queiroz
authors.forename: Selma Andrade
authors.initials: SA
authors.lastname: Gomes
authors.forename: Ana Carolina
authors.initials: AC
authors.lastname: Magalhães de Andrade Góes
authors.forename: Geraldo Rodrigues
authors.initials: GR
authors.lastname: Garcia Armoa
authors.forename: Carlos Humberto
authors.initials: CH
authors.lastname: Marques
authors.forename: Márcia Terezinha
authors.initials: MT
authors.lastname: Baroni de Moraes
mesh_terms: D000478:Alkylation; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D026481:Blotting, Far-Western; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010084:Oxidation-Reduction; D011498:Protein Precursors / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014759:Viral Envelope Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D006023:Glycoproteins; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C118669:S envelope protein, hepatitis B virus; D014759:Viral Envelope Proteins; C050806:presurface protein 1, hepatitis B surface antigen; C050807:presurface protein 2, hepatitis B surface antigen