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.