Purification of a Chlamydia trachomatis-specific antigen by immunoadsorption with monospecific antibody.
This study describes the isolation and partial characterization of a Chlamydia trachomatis specific antigen. A species-specific antigen of C. trachomatis (antigen-0.65) was identified by two-dimensional immunoelectrophoresis. Antiserum specific for antigen-0.65 was prepared in rabbits by immunizing with agarose-gel precipitates excised from two-dimensional immunoelectrophorograms. Purified gamma-globulins from antigen-0.65 specific serum were coupled to the N-hydroxysuccinimide ester derivative of agarose which was then used for the immunoadsorbent purification of antigen-0.65 from Triton X-100 solubilized lymphogranuloma venereum (L2/434/Bu) organisms. The isolated antigen was immunochemically pure when tested against rabbit antiserum prepared to LGV-434 organisms by using rocket and two-dimensional immunoelectrophoresis. Antigenicity was destroyed by protease treatment and heating at 56 degrees C for 30 min, but the antigen was stable to ribonuclease, deoxyribonuclease, periodate oxidation and pH extremes of 2.2 and 10.6. Polyacrylamide gel electrophoresis of purified antigen showed a major protein band with an apparent m.w. of 155,000.
D000818:Animals; D000907:Antibodies, Bacterial; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D002692:Chlamydia trachomatis / Q000276:immunology*; D002850:Chromatography, Gel; D000939:Epitopes; D000431:Ethanol / Q000494:pharmacology; D005026:Ethylene Glycols / Q000494:pharmacology; D007122:Immunoelectrophoresis; D007163:Immunosorbent Techniques; D008219:Lymphogranuloma Venereum / Q000276:immunology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011817:Rabbits
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D000939:Epitopes; D005026:Ethylene Glycols; D000431:Ethanol; D011092:Polyethylene Glycols