Acellular pertussis vaccine toxoided with carbodiimide

[摘要] Pertussis toxin (PT) and filamentous haemagglutinin (FHa) were extracted from the culture fluid of Bo pertussis grown in (1) modified Stainer and Scholte liquid medium (SS-X), static, (2) SS-X medium plus cyclodextrin, shaken, (3) eyelodextrin-liquid (CL) medium, shaken, by a single-step procedure using dye-ligand affinity chromatography. The addition of cyclodextrin to SS-X medium and aeration greatly enhanced the amount of PT and FHA extracted with Blue Sepharose, and the resulting PT and FHA ratio in the antigen preparation depended on the cultural conditions. With cyclodextrin-supplemented media (SS-X, CL) and aeration, it appeared that the total protein extracted was accounted for as PT and FHA, whereas with SS-X (static), only approximately 507o (w/w) was accounted for as both antigens. Culture of B. pertussis in SS-X medium supplemented with cyclodextrin (shaken), yielded the most PT, whereas growth in the CL medium (shaken) yielded the most FHa, Minor protein components were detected in many of the PT-FHa preparations, and endotoxin was present to 1-27, (w/w) of total protein in antigen preparations AP-16 and AP-17. The mixture of PT and FHa was toxoided with a water-soluble carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, HCl (EDAC), The toxoiding conditions most suitable for elimination of the pathophysiological activities of the A-protomer and B-oligomer of PT were reaction of EDAC with protein at a ratio of 80:1 by weight, at 37

[发布机构] University:University of Glasgow

[关键词] Immunology