RECOMBINANT TECHNOLOGY IN THE PREPARATION OF IMMUNOGEN AND ENZYMATIC TRACER - APPLICATION TO THE DEVELOPMENT OF AN ENZYME-IMMUNOASSAY FOR RAT PROLACTIN

被引：6

作者：

EZAN, E ^{[1
]}

DUCANCEL, F ^{[1
]}

GILLET, D ^{[1
]}

DREVET, P ^{[1
]}

MENEZ, A ^{[1
]}

GROGNET, JM ^{[1
]}

BOULAIN, JC ^{[1
]}

机构：

[1] CTR ETUD SACLAY, CEA, DEPT INGN & ETUD PROT, F-91191 GIF SUR YVETTE, FRANCE

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 1994年 / 169卷 / 02期

关键词：

RAT PROLACTIN; ENZYME IMMUNOASSAY; ALKALINE PHOSPHATASE; PLASMA; RECOMBINANT TECHNOLOGY;

D O I：

10.1016/0022-1759(94)90264-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A competitive enzyme immunoassay for rat prolactin using an immunogen and a tracer obtained by recombinant DNA technology is described. Polyclonal antibodies were raised in rabbits immunized with a purified chimeric protein consisting of rat prolactin fused with two synthetic immunoglobulin G binding domains derived from staphylococcal Protein A. The enzymatic tracer was obtained using an expression system which permits insertion of rPRL DNA sequence in the alkaline phophatase gene. Antibodies and tracer were used to develop a solid-phase competitive immunoassay for the measurement of rat prolactin in plasma with a minimal detectable concentration of 0.5 ng/ml. The mean intra- and inter-assay coefficients of variation were 7.8 and 13.2%, respectively. Rat plasma concentrations measured with this assay correlated well with those obtained with a conventional enzyme immunoassay (r = 0.993, slope = 1.037, n = 24).

引用

页码：205 / 211

页数：7

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