Identification of leptospiral flagellar antigens by gel electrophoresis and immunoblotting

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Abstract
Summary. Flagella extracted from five serovars, representative of the pathogenic and saprophytic species of the Leptospiraceae, were morphologically similar. Analysis of Leptospira interrogans flagellar preparations by polyacrylamide gel electrophoresis revealed three common major bands in the (30-40) x 103-mol. wt region, and serovar-specific bands in the lower region of the gels. Although some differences were observed, flagella extracted from L. biflexa serovar patoc and Leptonema illini revealed similar electrophoretic profiles to those seen in L. interrogans flagella. Immunoblot analysis showed that while flagellar components in the (20-30) x 103-mol. wt region were recognised only by homologous rabbit antisera, a major protein doublet of (33-34) x 103 or (35-36) x 103-mol. wt, depending on the species, was also demonstrated by heterologous antisera. The serovar-specific bands in the (20-30) x 103-mol. wt region were composed of lipopolysaccharide (LPS). These results show that leptospiral flagella are immunogenic and contain antigens which are conserved among the different genera of the family Leptospiraceae.