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.
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.
Alves Vianna
Carlos Otávio
CO
Laboratory of Monoclonal Antibodies Technology, Bio-Manguinhos, Oswaldo Cruz Foundation, FIOCRUZ, P.O. Box 926, Rio de Janeiro, Brazil.
da Silva E Mouta Júnior
Sérgio
S
da Glória Teixeira Martins
Maria
M
Batoreu
Nadia Maria
NM
Queiroz
João Luiz
JL
Gomes
Selma Andrade
SA
Magalhães de Andrade Góes
Ana Carolina
AC
Garcia Armoa
Geraldo Rodrigues
GR
Marques
Carlos Humberto
CH
Baroni de Moraes
Márcia Terezinha
MT
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
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