Evaluation of Filamentous Hemagglutinin in Combination with other Bordetella pertussis Antigens as Acellular Vaccine against Whooping Cough

Abstract

B. pertussis strain 134 was grown in a large fermentor according to standardized procedures for preparation of the whole cell vaccine. The culture supernatant was collected, clarified by filtration and concentrated by ultrafiltration. Purified PT and FHA were prepared by chromatography of the concentrate successively on butyl and fetuin columns. Antisera to the purified pertussis antigens were prepared in rabbits. The recovered antigens are highly purified as judged by SDS­ pAGE and immunoblots of the crude purified fractions The FHA and PT content of the Sephacryl fractions were determined in ELISA and the integrity of the PT content was determined in a functional cell culture assay.

Fractions containing different concentrations of the FHA and PT were prepared from the concentrated filtrate by gel filtration chromatography using Sephacryl S-300. The protective immunogenicity of the fractions was determined in vaccination challenge experiments "the IC mouse assay ". Mice immunized with the glutaraldehyde inactivated Sephacryl fractions were challenged with a virulent B. pertussis strain, and the potency of each fraction was calculated according to a