The heat-labile toxin (HLT) of Bordetella pertussis was purified by column chromatography on DEAE-cellulose, salt fractionation and preparative acrylamide gel electrophoresis.

A cell wall fraction obtained by sonic oscillation, yielded a Bordetella pertussis O antigen which possessed the ability to both adsorb O agglutinins and to produce the O agglutinins in animals. It was purified by a phenol water extraction and centrifugation method. Physico‐chemical and biological properties of the O antigen were similar to that of the so‐called endotoxic lipopolysaccharide. It contained 2.6% nitrogen, 1.2–1.5% phosphorus, 9.0–11.0% heptose, 0.3% 2‐keto‐3‐deoxyoctonate (KDO), 13.3–13.4% as total carbohydrate, 16–16.5% hexosamine and 25–32% lipid. Homogeneity of the purified O antigen was confirmed by ultracentrifugation, electronmicrograph and gel‐diffusion tests. It possessed significant pyrogenicity, adjuvant effect on antibody production, and toxicity to animals, but not protective antigenicity.

Relationship between K agglutinogenic factors of Bordetella pertussis were analyzed by agglutinin-adsorption tests. Factors 1 and 3 corresponded to L agglutinogen, while 2 and 4 corresponded to S agglutinogen. The experiment also indicates that B. pertussis phase I strains belong to a single serotype, while types 1-2-3, 1-2, 1-3 and 1 strains belong to the author's intermediate phase which probably arose from phase I parents. Strains isolated by us in 1952 to 1953 were type 1-2-3-4 which possessed both L and S, but a few strains isolated in 1962 and 1966 were type 1-3. The occurrence of such a defect in antigens was discussed. A single phase I vaccine seemed to give enough protection against various serotypes of pertussis organisms.

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