Harding G B, Doyle R J, Westphal U
Mol Immunol. 1982 Feb;19(2):267-79. doi: 10.1016/0161-5890(82)90340-6.
Antibodies against progesterone were induced in guinea pigs of both sexes by injection of progesterone-y beta-hemisuccinate conjugated to bovine serum albumin (BSA) in a ratio of 16 moles of steroid per mole of protein. The concentration of antibody binding sites for progesterone of the animals studied ranged from 5 to 20 microM. The expected heterogeneity of binding affinity for progesterone was observed with two major populations apparently predominating. On bound progesterone with an average affinity greater than 2 X 10(9) M-1 and the other showed an average affinity less than or equal to 6 X 10(6) M-1. The antibodies were fond to be stable to extremes of pH and temperature in serum as well as in solutions of ammonium sulfate precipitates. The antibodies were not stable, however, in a more highly purified form. Attempts to obtain active preparations in high yield by purification beyond the ammonium sulfate step were unsuccessful. Competition studies and direct analysis with radiolabeled steroids showed the high-affinity population to be relatively specific for progesterone binding, whereas other steroids were bound according to the polarity rule indicating that the binding forces are predominantly hydrophobic.
通过以每摩尔蛋白质16摩尔类固醇的比例注射与牛血清白蛋白（BSA）偶联的孕酮 -γ-半琥珀酸酯，在雄性和雌性豚鼠中诱导出抗孕酮抗体。所研究动物的孕酮抗体结合位点浓度范围为5至20微摩尔。观察到孕酮结合亲和力预期的异质性，两个主要群体显然占主导地位。一个群体对结合孕酮的平均亲和力大于2×10⁹ M⁻¹，另一个群体的平均亲和力小于或等于6×10⁶ M⁻¹。这些抗体在血清以及硫酸铵沉淀溶液中对极端pH和温度都很稳定。然而，抗体以更高纯度形式时不稳定。试图通过超出硫酸铵步骤的纯化来高产获得活性制剂未成功。竞争研究和用放射性标记类固醇的直接分析表明，高亲和力群体对孕酮结合相对特异，而其他类固醇根据极性规则结合，表明结合力主要是疏水性的。
