Studies on the purification of chlamydial agents grown in yolk sacs of embryonated eggs using disulphide-linked immunosorbents and enzymes.
J Gen Microbiol, 1975/7;89(1):11-25.
PMID: 239096
Abstract
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.
MeSH terms
Adsorption; Animals; Antibodies, Bacterial; Antibody Formation; Antigen-Antibody Complex; Antigen-Antibody Reactions; Antigens; Antigens, Bacterial; Cell Line; Chick Embryo; Chlamydia; Disulfides; Female; HeLa Cells; Hydrogen-Ion Concentration; Immune Sera; Immunodiffusion; Immunologic Techniques; Phospholipases; Trypsin; Vitelline Membrane
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