A sensitive, non-isotopic immunoassay for progesterone using the avidin-biotin system

被引:3
作者
Kanzaki, M [1 ]
Iwasawa, A [1 ]
机构
[1] GIFU UNIV,FAC AGR,DEPT ANIM SCI & TECHNOL,GIFU 50111,JAPAN
来源
BIOMEDICAL RESEARCH-TOKYO | 1995年 / 16卷 / 06期
关键词
D O I
10.2220/biomedres.16.381
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
A non-isotopic immunoassay for progesterone has been developed using streptavidin-biotin system and time-resolved fluorometry. A biotinylated progesterone derivative and a progesterone standard or sample was allowed to react competitively with rabbit anti-progesterone in microtiter wells coated with anti-rabbit gamma-globulin (second antibody). A fluorescent product was then generated on the solid-phase by adding streptavidin labeled with europium (Eu, a lantanide element). The fluorescent complex, (second antibody)-(anti-progesterone)-(biotinylated progesterone derivative)-(Eu-labeled streptavidin) was quantified using a time-resolved fluorometer. This assay for progesterone was highly sensitive, precise and accurate. The lower detection limit was 5 pg/ml. The intra-assay coefficients of variation (CV) were 3.70-5.20% and the inter-assay CV were 7.31-8.23%. The correlation with radioimmunoassay was good (r = 0.965), This assay was used to measure the amount of progesterone that accumulated in the culture medium of rat granulosa cells. The accumulation increased with the amount of insulin-like growth factor-I added to the culture in the presence of follicle-stimulating hormone. The present assay with streptavidin-biotin system can be applied to various tracers such as enzymes and fluorescent materials. This approach should also be widely applicable to antigens and haptens that can be labeled with biotin.
引用
收藏
页码:381 / 386
页数:6
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