Purification and characterisation of a fimbrial haemagglutinin from Bordetella pertussis for use in an enzyme-linked immunosorbent assay.
The fimbrial haemagglutinin (F-HA) of Bordetella pertussis grown on solid medium was extracted with 1M sodium acetate for 72 h at 20 degree C, and partially purified by Sephacryl S-300 gel chromatography. A pooled fraction with fimbrial haemmagglutinating activity was shown to contain fimbriae 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 measureable 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.
D000907:Antibodies, Bacterial / Q000032:analysis; D001886:Bordetella pertussis / Q000276:immunology*; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006388:Hemagglutinins / Q000302:isolation & purification*; D006801:Humans; D014917:Whooping Cough / Q000276:immunology
D000907:Antibodies, Bacterial; D006388:Hemagglutinins