Purification of nonlipopolysaccharide antigen from Brucella abortus during preparation of antigen used for indirect hemolysis test | Zendy

E. M. Hoffmann | Zendy; Jeri Joan Houle | Zendy

Open Access

Purification of nonlipopolysaccharide antigen from Brucella abortus during preparation of antigen used for indirect hemolysis test

Author(s) -

E. M. Hoffmann,

Jeri Joan Houle

Publication year - 1986

Publication title -

journal of clinical microbiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.349

H-Index - 255

eISSN - 1070-633X

pISSN - 0095-1137

DOI - 10.1128/jcm.24.5.779-784.1986

Subject(s) - complement fixation test , antigen , hemolysis , brucella , chromatography , polyacrylamide gel electrophoresis , brucellaceae , size exclusion chromatography , biology , microbiology and biotechnology , antibody , chemistry , brucella abortus , brucellosis , biochemistry , brucella melitensis , virology , serology , immunology , enzyme

The indirect hemolysis test (IHLT) for the diagnosis of brucellosis uses a lipopolysaccharide (LPS) antigen obtained by dimethyl sulfoxide extraction of Brucella abortus. We showed that a non-LPS antigen can be obtained as a by-product of the IHLT antigen preparation. The antigen was purified to homogeneity by a combination of gel-filtration chromatography and ion-exchange chromatography. The substance contained 8% protein and about 65% carbohydrate. The molecular weight of the primary unit was 19,750, when analyzed by polyacrylamide gel electrophoresis under denaturing conditions. The non-LPS antigen, which is serologically identical to B. abortus smooth LPS O antigen, did not bind to cell membranes. However, it could be used to detect specific antibodies by complement fixation, precipitation in agarose gels, and inhibition of the IHLT.

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