Purification and Characterisation of a Fimbrial Haemagglutinin from Bordetellapertussis for Use in an Enzyme-linked Immunosorbent Assay

1 Departments of Production and Bacteriology Stockholm, Sweden 2 National Bacteriological Laboratory, S-105 21 Stockholm, Sweden Received April 10, 1981 Accepted July 24, 1981 The fimbrial haemagglutinin (F-HA) of Bordetella pertussis grown on solid medium was extracted with 1m sodium acetate for 72 h at 20°C, and partially purified by Sephacryl S-300 gel chromatography. A pooled fraction with fimbrial haemagglutinating activity was shown to contain fimbriae of the expected morphology by electron microscopy. Chemical and biological assays showed that the F-HA fraction contained some heat-labile agglutinogen and lipopolysaccharide but no measurable lymphocytosis-promoting factor or heat-labile toxin. The F-HA fraction used as antigen in an enzyme-linked immunosorbent assay (ELISA) permitted the detection of antibodies in convalescent serum from a patient with whooping cough. The impurities, heat-labile agglutinogens and lipopolysaccharide, did not contribute to the ELISA activity. The method for preparation of the F-HA antigen is simple, reproducible and gives a high yield.