牛血浆中生长激素的生物素-抗生物素蛋白夹心酶联免疫吸附测定

A biotin-avidin sandwich enzyme-linked immunosorbent assay of growth hormone in bovine plasma.

作者信息

Secchi C, Biondi P A, Berrini A, Simonic T, Ronchi S

机构信息

Institute of Veterinary Physiology and Biochemistry, University of Milan, Italy.

出版信息

J Immunol Methods. 1988 May 25;110(1):123-8. doi: 10.1016/0022-1759(88)90091-9.

DOI:10.1016/0022-1759(88)90091-9

PMID:3373001

Abstract

Purified pituitary bovine growth hormone (bGH) has been used to develop a homologous sandwich enzyme-linked immunosorbent assay in which affinity-purified antibodies are immobilized on microtiter plates. Bovine GH bound to specific antibody is then revealed with a second anti-bGH antibody labeled with biotin and peroxidase-conjugated avidin. The method requires only 48 h, including the coating step, and has a sensitivity as low as 0.25 ng/ml of bGH. Statistical analyses (test of parallelism, cross-reactivity among bGH and GH of various species and bovine prolactin, recovery test, within- and between-assay variation, comparison with radioimmunoassay) confirm the high specificity and reproducibility of the method.

摘要

纯化的牛垂体生长激素（bGH）已被用于开发一种同源夹心酶联免疫吸附测定法，其中亲和纯化的抗体固定在微量滴定板上。与特异性抗体结合的牛生长激素随后用生物素标记的第二抗bGH抗体和过氧化物酶偶联的抗生物素蛋白进行检测。该方法仅需48小时，包括包被步骤，对bGH的灵敏度低至0.25 ng/ml。统计分析（平行性测试、bGH与各种物种的生长激素及牛催乳素之间的交叉反应性、回收率测试、批内和批间变异、与放射免疫测定法的比较）证实了该方法具有高特异性和可重复性。