Purification of Treponema pallidum, Nichols strain, by two-step column chromatography.
A rapid and simple purification method for Treponema pallidum, Nichols strain, the etiological agent of venereal syphilis, was developed. A 40-ml suspension of organisms (1.10(9)/ml) was extracted from rabbit testicular tissue and solubilized with a non-ionic detergent, 1-O-n-octyl-beta-D-glucopyranoside. Solubilized antigens were purified by cation-exchange and hydroxyapatite column chromatography. The overall recovery of immunoreactive material was 48.3% and the specific activity increased. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting analysis confirmed the purity and species specificity of the purified antigen.
Matsumoto
M
M
Sekisui Chemical Co., Ltd., Medical Research Laboratory, Osaka, Japan.
Ishikawa
F
F
D000818:Animals; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange; D003902:Detergents; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007202:Indicators and Reagents; D008297:Male; D011817:Rabbits; D013737:Testis / Q000382:microbiology*; D014210:Treponema pallidum / Q000276:immunology*
D000942:Antigens, Bacterial; D003902:Detergents; D007202:Indicators and Reagents; D017886:Durapatite