Clinical Chemistry Applied Biosystems
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Clinical Chemistry 19: 1027-1030, 1973;
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Clinical Chemistry, Vol 19, 1027-1030, Copyright © 1973 by the American Association for Clinical Chemistry

Purification of Progesterone Antiserum for Radioimmunoassay, by Chromatography on QAE-Sephadex A-50

Frank H. Bodley 1, Alcide Chapdelaine 1, and Kenneth D. Roberts 1

1 Departments of Biochemistry and of Medicine, University of Montreal and Maisonneuve-Rosemont Hospital, Montreal, Quebec, Canada.

Antibodies were generated in sheep after administration of the antigen, progesterone-11agr-succinyl-bovine serum albumin. This antiserum was purified 10-fold by treatment with "Aerosil" and column chromatography on "QAE-Sephadex A-50." In addition to an increased specific activity (cpm progesterone bound/g of protein) the purified antibody fraction was less cross-reactive with pregnenolone and desoxycorticosterone than was the crude antiserum. A sensitivity of 10 pg of progesterone was obtained with the purified antibody preparation, compared to 25 pg for the crude antiserum. This method of purification of antisera was devised in order to concentrate the antibody fraction before covalent coupling to insoluble supports such as activated arylamine-glass particles.

Submitted on May 18, 1973
Accepted on June 2, 1973






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Copyright © 1973 by the American Association for Clinical Chemistry.