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针对孕酮的高亲和力单克隆抗体的制备。

The production of high affinity monoclonal antibodies to progesterone.

作者信息

Fantl V E, Wang D Y, Knyba R E

出版信息

J Steroid Biochem. 1982 Aug;17(2):125-30. doi: 10.1016/0022-4731(82)90110-8.

Abstract

Using different immunisation regimens and the antigens 6 beta-hydroxyprogesterone hemisuccinate and 11 alpha-hydroxyprogesterone hemisuccinate conjugated to bovine serum albumin, we have produced 35 monoclonal antibodies against progesterone with a wide range of specificities and affinities (Ka = 8 x 10(7)-3 x 10(10) M-1). The 10 antibodies raised against the 6 beta-antigen showed no advantage in terms of specificity when compared with those raised against the 11 alpha-antigen. The four antibodies with the best binding characteristics were all derived from fusion experiments performed after long (5 booster injections) immunisation with the 11 alpha-antigen. Although the specificities of these monoclonal antibodies were similar to those of a high affinity rabbit antiserum raised against the same antigen, a measurable improvement in sensitivity was obtained when one of these four antibodies was used for radioimmunoassay.

摘要

我们使用不同的免疫方案以及与牛血清白蛋白偶联的6β-羟基孕酮半琥珀酸酯和11α-羟基孕酮半琥珀酸酯抗原,制备了35种针对孕酮的单克隆抗体,这些抗体具有广泛的特异性和亲和力(Ka = 8×10⁷ - 3×10¹⁰ M⁻¹)。与针对11α-抗原产生的抗体相比,针对6β-抗原产生的10种抗体在特异性方面没有优势。具有最佳结合特性的四种抗体均来自用11α-抗原进行长期(5次加强注射)免疫后进行的融合实验。尽管这些单克隆抗体的特异性与针对相同抗原产生的高亲和力兔抗血清的特异性相似,但当使用这四种抗体中的一种进行放射免疫测定时,灵敏度得到了可测量的提高。

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