Purification of genus-specific chlamydial antigen and its separation into several components by ion-exchange chromatography.
Sodium deoxycholate-extracted genus-specific chlamydial antigen was purified from contaminating substances by ion-exchange chromatography with DEAE-sephacel, resulting in a decrease in the complement-fixing activity of the antigen, whereas the enzyme-linked immunosorbent assay activity increased. By successive elution with 0.5, 1.0, and 1.5 M acetate buffer at least three clearly separated components were consistently recovered in 14 trials. The identity of these components as genus-specific chlamydial antigen was demonstrated in enzyme-linked immunosorbent assay tests with specific antisera. The antigenic activity of these components was not diminished by prior treatment of the chlamydial particles with pronase. Antiserum prepared by immunization of rabbits with the antigenic component I of an egg-propagated antigen reacted predominantly with the antigenic components I and II of a cell culture-propagated antigen.
D000956:Antigens, Viral / Q000302:isolation & purification*; D002689:Chlamydia / Q000276:immunology*; D002852:Chromatography, Ion Exchange; D003168:Complement Fixation Tests; D003840:Deoxycholic Acid / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D011402:Pronase / Q000494:pharmacology
D000956:Antigens, Viral; D003840:Deoxycholic Acid; D011402:Pronase