Open AccessPurification and Characterisation of a Fimbrial Haemagglutinin from Bordetellapertussis for Use in an Enzyme-linked Immunosorbent Assay
Author(s) -
Per Askelöf,
Marta Granström,
Pekka Gillenius,
Annelie Lindberg
Publication year - 1982
Publication title -
journal of medical microbiology/journal of medical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.91
H-Index - 117
eISSN - 1473-5644
pISSN - 0022-2615
DOI - 10.1099/00222615-15-1-73
Subject(s) - bordetella pertussis , enzyme , fimbria , antigen , chemistry , microbiology and biotechnology , toxin , fraction (chemistry) , antibody , lipopolysaccharide , chromatography , bacteria , biology , biochemistry , escherichia coli , immunology , genetics , gene
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.