title: Establishment and validation of an ELISA for the quantitation of HBsAg in recombinant hepatitis B vaccines.
abstract: Commercial enzyme-linked immunosorbent assay (ELISA) kits for the determination of the in vitro potency of recombinant hepatitis B vaccines, which detect hepatitis B surface antigen (HBsAg), have been used frequently as an alternative for traditional in vivo potency tests. With the constant need for validation procedures, an ELISA that could be employed to determine the in vitro potency of five recombinant hepatitis B vaccines simultaneously was established using two monoclonal antibodies. The use of two monoclonal antibodies produced "in house" specific for the small envelope protein S of the hepatitis B virus (HBV) resulted in the production of a highly specific, sensitive and stable ELISA. The standard ELISA parameters used in this study, considering the HBsAg content of each recombinant hepatitis B vaccine evaluated, resulted in a standard curve that could be applied for potency evaluations of different, commercial hepatitis B vaccine lots.
authors.affiliation: Department of Immunology, National Institute of Quality Control in Health (INCQs), Oswaldo Cruz Foundation (FIOCRUZ), P.O. Box 926, Rio de Janeiro, Brazil.
authors.forename: Catia Inês
authors.initials: CI
authors.lastname: Costa
authors.forename: Isabella Fernandes
authors.initials: IF
authors.lastname: Delgado
authors.forename: Jaline Alves Cabral
authors.initials: JA
authors.lastname: da Costa
authors.forename: Renata Faria
authors.initials: RF
authors.lastname: de Carvalho
authors.forename: Sérgio da Silva
authors.initials: Sda S
authors.lastname: Mouta
authors.forename: Carlos Otávio Alves
authors.initials: CO
authors.lastname: Vianna
authors.forename: Márcia Terezinha Baroni
authors.initials: MT
authors.lastname: de Moraes
mesh_terms: D000911:Antibodies, Monoclonal; D057205:Automation, Laboratory; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards*; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens / Q000032:analysis*; D017325:Hepatitis B Vaccines / Q000032:analysis*; D006801:Humans; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014614:Vaccines, Synthetic / Q000032:analysis*
chemical_list: D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D011933:Reagent Kits, Diagnostic; D014614:Vaccines, Synthetic