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Purification of Semliki Forest Virus Using Disulphide-linked Immunosorbents Derived from Rabbit or Fowl Antibody

Department of Microbiology, University of Birmingham, Birmingham 15

Semliki Forest virus in the crude supernatant from a culture of chick embryo cells was specifically adsorbed by a disulphide-linked immunosorbent prepared from the -globulins of rabbits immunized by Semliki Forest virus grown in mouse brain. Two to 6 ml. of rabbit antiserum of neutralization index 4 to 5 yielded 10 mg. of immunosorbent which bound 1 to 6 x 10¹⁰ p.f.u. from 2 to 30 ml. of culture supernatant. Virus, free from most chick cell constituents, could be eluted from the immunosorbents by treatment with bicarbonate/carbonate buffer at pH 11.1. The immunosorbents could be cycled at least five times and about 50% of the adsorbed virus infectivity recovered at each cycle after the first, from which only 10 to 20% was recovered. In capacity experiments, immunosorbents derived from chicken antisera against Semliki Forest virus grown in chick cells showed no advantages over those derived from rabbit antisera.

Received 23 July 1969; accepted 2 October 1969.