1994, Number 2
Production of antibodies against progesterone from the eggyolk of hens and from rabbit blood serum to be used in radioimmunoassay
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ABSTRACTLeghorn hens and New Zealand rabbits were immunized against progesterone, in order to produce antibodies to be used in radioinmunoassay (RIA). Acetone and 2-propanol were used to extract antibodies from the yolk of eggs of immunized hens. Antibody extraction from rabbit blood serum was done by precipitation with amonium sulfate. Antibodies from egg yolk had high affinity and specificity for progesterone. Sensitivity of the RIA established with these antibodies, was 0.27 ng/ml, with intra- and interassay coefficients of variation of 19.6 and 22.2%, respectively. There was a significant correlation (0.94) between the progesterone concentrations determined with egg-yolk antibodies and those found using a commercial kit. Antibodies from rabbit serum had also high affinity and specificity for progesterone. RIA, established with these antibodies, had a sensitivity of 0.23 ng/ml, with intra- and interassay coefflcients of variation of 14.7 and 22.7%, respectively. The correlation between progesterone values determined with rabbit antiserum and those obtained with a commercial kit was 0.96. The yield of antibodies from egg yolk was 2.5 times higher than that of rabbit serum. It is concluded, that egg yolk of immunized hens is a practical source of antiprogesterone antibodies with similar quality to that of antibodies extracted from the blood serum of immunized rabbits.
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