用于白细胞介素-3时间分辨荧光免疫测定的铕-链霉亲和素的制备

Preparation of europium-streptavidin in a time-resolved fluoroimmunoassay for interleukin-3.

作者信息

Knopf H P, Papoian R

机构信息

Department of Biopharmaceutics, Sandoz Pharma, Basel, Switzerland.

出版信息

J Immunol Methods. 1991 Apr 25;138(2):233-6. doi: 10.1016/0022-1759(91)90171-b.

DOI:10.1016/0022-1759(91)90171-b

PMID:2033276

Abstract

The preparation and immunoassay performance of europium-labeled streptavidin is described. The Eu-streptavidin conjugate can serve as a general detection reagent in time-resolved fluoroimmunoassays (TRFIA). The usefulness of such a strategy has previously been demonstrated with the Eu3+ chelate 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (Diamandis et al. 1988). In this report the conjugation of streptavidin was accomplished with the Eu3+ chelate of N1-(p-isothiocyanatobenzyl)diethylenetriamine-N1,N3,N3(+)-tetraace tic acid, using a 50 M excess of the label. The conjugation ratio of Eu3+/streptavidin was 16. The use of the Eu-streptavidin reagent in a two-site immunometric assay to measure human recombinant interleukin-3 in human plasma, showed that the useful range of the assay was 20-25,000 pg/ml.

摘要

本文描述了铕标记链霉亲和素的制备及其免疫分析性能。铕标记的链霉亲和素缀合物可作为时间分辨荧光免疫分析（TRFIA）中的通用检测试剂。此前已证明，采用铕离子螯合物4,7-双（氯磺苯基）-1,10-菲咯啉-2,9-二羧酸的这种策略是有用的（Diamandis等人，1988年）。在本报告中，链霉亲和素的缀合是用N1-（对异硫氰酸苄基）二乙烯三胺-N1,N3,N3（+）-四乙酸的铕离子螯合物完成的，标记物过量50倍。铕离子/链霉亲和素的缀合比为16。在两点免疫分析中使用铕标记链霉亲和素试剂检测人血浆中的人重组白细胞介素-3，结果表明该分析的有效范围为20 - 25,000 pg/ml。