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通过单克隆抗体定位的人胎盘碱性磷酸酶和睾丸胎盘样碱性磷酸酶的抗原决定簇

Antigenic determinants of human placental and testicular placental-like alkaline phosphatases as mapped by monoclonal antibodies.

作者信息

Millán J L, Stigbrand T

出版信息

Eur J Biochem. 1983 Oct 17;136(1):1-7. doi: 10.1111/j.1432-1033.1983.tb07697.x.

Abstract

Placental alkaline phosphatase (PLAP) in humans shows a high degree of genetic polymorphism as disclosed by electrophoretic analysis. Human testes contain trace amounts of a PLAP-like enzyme, that although immunologically cross-reactive with PLAP, shows unique catalytic properties. As an alternative approach to study enzyme polymorphism we have developed monoclonal antibodies to purified allelic variants of PLAP. Five different monoclonal antibodies are described in this report. The antibodies react with different epitopes on the PLAP molecule. Both conformational dependent and independent determinants are detected. Two epitopes are modified when comparing the S and F allelic variants of PLAP. One epitope is common to PLAP and the intestinal isoenzyme of alkaline phosphatase. The five epitopes appear to be mapped on two rather distant antigenic domains. Combinations of any two antibodies binding to different domains give immunoprecipitates with PLAP on Ouchterlony tests and give good response in sandwich enzyme-linked immunosorbent assays. A study of PLAP-like enzyme in 32 individual testis samples indicates differences in four of the epitopes when compared with PLAP. Four types of testicular enzymes can be distinguished based on their reactivities. These results indicate structural differences between the testicular PLAP-like enzyme and PLAP. These differences are compatible with an underlying genetic mechanism.

摘要

电泳分析显示,人胎盘碱性磷酸酶(PLAP)具有高度的遗传多态性。人类睾丸含有微量的一种PLAP样酶,该酶虽然与PLAP存在免疫交叉反应,但具有独特的催化特性。作为研究酶多态性的另一种方法,我们制备了针对PLAP纯化等位基因变体的单克隆抗体。本报告描述了五种不同的单克隆抗体。这些抗体与PLAP分子上的不同表位发生反应。检测到了构象依赖性和非依赖性决定簇。比较PLAP的S和F等位基因变体时,有两个表位发生了改变。一个表位是PLAP和碱性磷酸酶肠型同工酶所共有的。这五个表位似乎定位在两个相距较远的抗原结构域上。在双向免疫扩散试验中,任何两种结合不同结构域的抗体组合都能与PLAP形成免疫沉淀物,并且在夹心酶联免疫吸附试验中表现出良好的反应。对32个个体睾丸样本中PLAP样酶的研究表明,与PLAP相比,其中四个表位存在差异。根据其反应性可区分出四种类型的睾丸酶。这些结果表明睾丸PLAP样酶与PLAP之间存在结构差异。这些差异与潜在的遗传机制相符。

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