Studies on the Purification of Chlamydial Agents Grown in Yolk Sacs of Embryonated Eggs Using Disulphide-linked Immunosorbents and Enzymes

Abstract

SUMMARYImmunosorbents 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.