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人绒毛膜促性腺激素β亚基核心片段的放射免疫测定法。

A radioimmunoassay for the core fragment of the human chorionic gonadotropin beta-subunit.

作者信息

Akar A H, Wehmann R E, Blithe D L, Blacker C, Nisula B C

机构信息

Developmental Endocrinology Branch, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.

出版信息

J Clin Endocrinol Metab. 1988 Mar;66(3):538-45. doi: 10.1210/jcem-66-3-538.

Abstract

We developed a RIA for the beta-core fragment of hCG that is excreted in the urine of pregnant women and some patients with cancer. We purified beta-core from crude commercial preparations of hCG (of which beta-core is a major constituent) derived from pregnancy urine and used this purified beta-core material to immunize rabbits. One antiserum (RW25) was particularly useful in that a RIA with purified beta-core as both radioligand and reference preparation had high sensitivity for beta-core detection and low cross-reactivity with other hCG-related molecules and glycoprotein hormones. The cross-reactivities of purified hCG (CR125), hCG beta (CR125), and hCG alpha (CR125) preparations were in each instance less than 3 x 10(-3) (wt/wt). The cross-reactivities of purified pituitary glycoprotein hormones were in each instance less than 2 x 10(-4) (wt/wt). Using the RW25 RIA, virtually all beta-core immunoreactivity in pregnancy urine eluted from Sephadex G-100 in a position coincident with that of purified beta-core. Urine from men and nonpregnant women contained very low levels of beta-core immunoreactivity (less than 6.5 micrograms/L), while urine from pregnant women and patients with testicular cancer or other neoplasms had levels of beta-core immunoreactivity ranging as high as 26,000 micrograms/L. We conclude that the improved specificity of our beta-core RIA will facilitate studies of the physiology and cancer biology of beta-core molecules.

摘要

我们开发了一种用于检测人绒毛膜促性腺激素(hCG)β核心片段的放射免疫分析方法(RIA),该片段存在于孕妇和一些癌症患者的尿液中。我们从妊娠尿液来源的hCG粗制商业制剂(β核心是其主要成分)中纯化β核心,并使用这种纯化的β核心物质免疫兔子。一种抗血清(RW25)特别有用,因为以纯化的β核心作为放射性配体和参考制剂的RIA对β核心检测具有高灵敏度,并且与其他hCG相关分子和糖蛋白激素的交叉反应性低。纯化的hCG(CR125)、hCGβ(CR125)和hCGα(CR125)制剂的交叉反应性在每种情况下均小于3×10⁻³(重量/重量)。纯化的垂体糖蛋白激素的交叉反应性在每种情况下均小于2×10⁻⁴(重量/重量)。使用RW25 RIA,妊娠尿液中几乎所有的β核心免疫反应性从Sephadex G - 100洗脱的位置与纯化的β核心一致。男性和非孕妇的尿液中β核心免疫反应性水平非常低(小于6.5微克/升),而孕妇以及睾丸癌或其他肿瘤患者的尿液中β核心免疫反应性水平高达26,000微克/升。我们得出结论,我们的β核心RIA提高的特异性将有助于对β核心分子的生理学和癌症生物学进行研究。

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