Purification of Coxiella burnetii antigen by gel-chromatography for use in enzyme-linked immunosorbent assay (ELISA).
Cell wall antigens from Coxiella burnetti were extracted by heat, ultrasonication, phenol, SDS or sodium deoxycholate. These antigens were subsequently purified by gel-chromatography yielding molecular weights of 700 and 270 KD. Using gel-chromatographical purification of antigens extracted by heat, ultrasonication or phenol Coxiella antigens nearly devoid of contaminations could be obtained. Of these highly purified antigens heat extracted antigen was treated with proteases, lysozyme or metaperiodate exemplarily. These treatments did neither destroy nor change its antigenic activity, except for the treatment with metaperiodate reducing the antigenic activity significantly. In further experiments heat extracted and gel-chromatographically purified Coxiella antigen was used for testing bovine sera in ELISA.
Sting
R
R
Staatliches Tierärztliches Untersuchungsamt Stuttgart, Germany.
Westphal
K
K
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000302:isolation & purification; D002417:Cattle; D002473:Cell Wall / Q000276:immunology; D002850:Chromatography, Gel / Q000662:veterinary; D016997:Coxiella burnetii / Q000276:immunology* / Q000648:ultrastructure; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D000954:Antigens, Surface