Studies on the purification of chlamydial agents grown in yolk sacs of embryonated eggs using disulphide-linked immunosorbents and enzymes.
Immunosorbents were derived from avid and non-avid sera raised in rabbits to multiple or single injections of chlamydiae passaged once or three times in HeLa cells after routine passage in eggs. Egg-derived suspensions of chlamydiae required pretreatment before application to immunosorbent columns; this was most conveniently done by fractionation on Sepharose 4B. Immunosorbents derived from avid serum had greater capacity than those from non-avid sera. However, organisms were desorbed in low yield from an avid immunosorbent, but in higher yields from non-avid immunosorbents. Even under the best conditions, the immunosorbents yielded suspensions of organisms still contaminated with egg antigens. Partially purified suspensions of chlamydiae were also treated with phospholipase C to yield suspensions of high purity.
D000327:Adsorption; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000917:Antibody Formation; D000936:Antigen-Antibody Complex; D000937:Antigen-Antibody Reactions; D000941:Antigens; D000942:Antigens, Bacterial; D002460:Cell Line; D002642:Chick Embryo; D002689:Chlamydia / Q000254:growth & development / Q000276:immunology / Q000302:isolation & purification*; D004220:Disulfides; D005260:Female; D006367:HeLa Cells; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D005779:Immunodiffusion; D007158:Immunologic Techniques; D010740:Phospholipases / Q000494:pharmacology*; D014357:Trypsin / Q000494:pharmacology; D014817:Vitelline Membrane / Q000276:immunology
D000907:Antibodies, Bacterial; D000936:Antigen-Antibody Complex; D000941:Antigens; D000942:Antigens, Bacterial; D004220:Disulfides; D007106:Immune Sera; D010740:Phospholipases; D014357:Trypsin