A BORDETELLA-PERTUSSIS ACELLULAR VACCINE CANDIDATE - ANTIGENIC CHARACTERIZATION AND ANTIBODY INDUCTION

A single-step chromatography on Matrex-Gel Blue A has been employed to obtain soluble extracts containing some of the most important antigen s of Bordetella pertussis, pertussis toxin (PT), filamentous hemagglut inin (FHA), pertactin (69-kDa outer membrane protein), fimbriae (FIM2 and FIM3) and adenylate cyclase (AC). Two supernatants, P19 (48.8 mg P T, 6.8 mg FHA, 17.3 mg AC, 13 mg FIM2 and 4.9 mg FIM3 per liter) and P 21 (0.1 mg PT, 0.07 mg FHA 0.46 mg FIM2 and 0.94 mg FIM3 per liter), r esulting from bacteria grown in Stainer-Scholte medium, were submitted to chromatography. Fractions with the antigens were obtained after st epwise elution with 60 mM sodium phosphate buffer, pH 6.0; 50 mM Tris- HCl, pH 7.4; 50 mM Tris-HCl, pH 7.4/0.75 M MgCl2; 50 mM Tris-HCl, pH 7 .4/4 M MgCl2 and 4 M urea. Preparations from P19 (containing 4.05 mu g PT, 8.14 mu g FHA, 6.3 mu g AC, 3.37 mu g 69-kDa, 9.54 mu g FIM2 and 2.23 mu g FIM3) and from P21 (with 0.175 mu g PT, 0.28 pg FHA, 0.002 m u g 69-kDa, 0.005 mu g FIM2 and 0.122 mu g FIM3) were detoxified with glutaraldehyde and tested as an acellular pertussis vaccine, These pro ducts were non-toxic for mice and induced high levels of antibodies ag ainst purified pertussis antigens, as judged by ELISA.