Production and Characterization of Antisera Against Steroids in Rabbits

Abstract

Antibodies against testosterone, progesterone and estradiol were raised by immunization of rabbits with testosterone-3-(o-carboxyrnethyl) oxime-bovine serum albumin (testosterone-3-BSA), progesterone-11-(o carboxymethyl) oxime-bovine serum albumin (progesterone 11-BSA), and estradiol-6-(0-carboxymethyl) oxime-bovine serum albumin (estradiol-6-BSA). The antisera have shown a high titer of 48-54% binding at the final dilution of 1:24,000. These antisera, therefore, have shown high affinity. The affinity: constant (Ka) calculated for testosterone, progesterone and estradiol antisera were 6.5x109 1/mol, 3.2x109 1/mol, and 0.29x109 1/mol, respectively. The specificity and cross-reactivity results have shown a high degree of specificity toward their respective homologous steroids. The mean percent recovery of unlabelled hormones added to samples was 105±10.63 for testosterone, 92.75±2.77 for progesterone and 107±5.38 for estradiol antisera. The sensitivity of testosterone, progesterone and estradiol standard of testosterone, progesterone and estradiol standard curves was 10,23 and picograms per 200 1, respectively, when calculated at 95% confidence limit.