title: Monoclonal antibody against non-dominant epitopes of HBV e Ag was raised by antigen-antibody co-immunization.
abstract: Detection of hepatitis B e antigen (HBeAg) in the sera of individuals infected with hepatitis B virus (HBV) can indicate both a high infectivity of the disease and a poor prognosis of disease treatment. Most of monoclonal antibodies raised against HBV e proteins interact with immuno-dominant epitopes, such as HBeAg-beta. In order to raise antibodies against non-dominant epitopes of HBV e protein, in this study, mice were immunized with both recombinant HBeAg (rHBeAg) and an anti-HBeAg antibody (EWB) recognizing a dominant antigenic epitope of HBeAg (HBeAg-beta epitope). With this strategy, we successfully selected two monoclonal antibodies, S-29-3 and S-72-3. Both S-29-3 and S-72-3 bind to recombinant HBeAg with a high affinity. The epitope mapping assay determined that the S-73-2 recognizes the N-terminal of HBeAg (1-118 aa) and the S-29-3 recognizes the C-terminal of HBeAg (91-149 aa). Further experiment showed that these two antibodies could be formed a pair-Abs that is used in detecting native HBeAg from the sera of HBV patients. The conclusion is that the developed method is useful to raise mAb against non-dominant epitopes in given Ag.
authors.affiliation: Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, Shanghai, PR China.
authors.forename: Ling Chen
authors.initials: LC
authors.lastname: Yan
authors.forename: Cai Ying
authors.initials: CY
authors.lastname: Jing
authors.forename: Chao Feng
authors.initials: CF
authors.lastname: Huang
authors.forename: Yong Yong
authors.initials: YY
authors.lastname: Ji
authors.forename: Gang
authors.initials: G
authors.lastname: Yao
authors.forename: Xing Feng
authors.initials: XF
authors.lastname: Cai
authors.forename: Bing
authors.initials: B
authors.lastname: Sun
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex; D000939:Epitopes / Q000276:immunology*; D006509:Hepatitis B / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D017325:Hepatitis B Vaccines; D006513:Hepatitis B e Antigens / Q000235:genetics / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D014765:Viral Vaccines
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000939:Epitopes; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D006513:Hepatitis B e Antigens; D014765:Viral Vaccines