Quantitation of adenylate cyclase of Bordetella pertussis by enzyme linked immunosorbent assay.
Bordetella pertussis produces extracytoplasmic adenylate cyclase toxin (AC) which has received considerable attention as a potential vaccine candidate. Great interest from laboratories involved in production, purification and quality control of acellular pertussis vaccine is focused on finding an appropriate technique for rapid and accurate quantitation of AC antigen. In this paper a competitive ELISA is proposed. A polystyrene microplate coated with purified AC was incubated with the sample to be tested plus anti-AC serum. The bound anti-AC antibodies were measured by sequential reaction with alkaline phosphatase-labelled anti-mouse IgG and p-nitrophenylphosphate. This method showed high specificity, with the 50% inhibition corresponding to 4 micrograms/ml of AC. It also proved to be useful to assess the presence of AC in culture supernatants, with high reproducibility.
Hozbor
D
D
Facultad de Ciencas Exactas, Universidad Nacional de La Plata, Argentina.
Chirdo
F G
FG
Rodriguez
M E
ME
Valverde
C
C
Yantorno
O
O
D000262:Adenylyl Cyclases / Q000032:analysis* / Q000276:immunology; D000818:Animals; D001886:Bordetella pertussis / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity
D000262:Adenylyl Cyclases