Characterization of a partially purified leptospiral genus-specific protein antigen.
A leptospiral genus-specific protein antigen (GP-Ag) was partially purified from Leptospira interrogans serovar kremastos strain Kyoto and canicola strain Hond Utrecht IV, by treating the organisms with Triton X-100 followed by a purification procedure including fractionation with DEAE-cellulose column chromatography and ethanol precipitation. Genus-specificity of the antigens from kremastos Kyoto and canicola was shown by the immunodiffusion (ID) test. Complement fixation (CF) test and enzyme-linked immunosorbent assay (ELISA) using GP-Ag from kremastos Kyoto showed the genus specificity of the GP-Ag. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of the antigens from kremastos Kyoto and canicola showed a major band with Coomassie blue stain. The band of GP-Ag from kremastos Kyoto was also shown by immunoprecipitation using the monoclonal antibody (GP-7) to GP-Ag from kremastos Kyoto. The molecular weight of the genus-specific antigen was estimated to be approximately 62 000. The genus specificity of the antigen was reduced by heating at 100 degrees C or treatment with proteolytic enzymes, but not reduced by treatment with sodium metaperiodate.
N
N
Sakamoto
E
E
Ono
H
H
Kida
R
R
Yanagawa
D000372:Agglutination Tests; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000032:analysis / Q000276:immunology*; D011232:Chemical Precipitation; D003168:Complement Fixation Tests; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006358:Hot Temperature; D005779:Immunodiffusion; D007921:Leptospira interrogans / Q000276:immunology*; D041341:Leptospira interrogans serovar canicola / Q000276:immunology*; D010504:Periodic Acid / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D000939:Epitopes; D010504:Periodic Acid; C009288:metaperiodate