Isolation and purification of a type-specific antigen from Chlamydia trachomatis propagated in cell culture utilizing molecular shift chromatography.
Various techniques have been utilized for antigen solubilization, isolation, and purification. This report is the first to describe the isolation and purification of a type-specific antigen from Chlamydia trachomatis serotype A grown in cell culture. The type-specific antigen was prepared from Chlamydia trachomatis serotype A organisms grown in baby hamster kidney cells (BHK21). The extraction process employed a combination of both pH change and Triton X-100 solubilization. The soluble extract was radioiodinated and subjected to ion exchange and gel filtration chromatography. The fractions eluted were tested for type specificity utilizing the IgG prepared from exhaustively cross-absorbed hyperimmune sera from rabbits immunized with homologous organisms. Molecular shift chromatography was employed for analysis. Small samples of the isolated antigen were later used as markers for preparation of larger quantities necessary for antigenic characterization. The purified type-specific antigen has a m.w. of 30,000 to 32,000.
D000818:Animals; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D002478:Cells, Cultured; D002690:Chlamydia Infections / Q000175:diagnosis; D002692:Chlamydia trachomatis / Q000276:immunology*; D002850:Chromatography, Gel; D006224:Cricetinae; D000939:Epitopes; D008970:Molecular Weight; D009500:Neutralization Tests; D011817:Rabbits
D000942:Antigens, Bacterial; D000939:Epitopes