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KMID : 0900919920160040325
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Korean journal of Animal Reproduction
1992 Volume.16 No. 4 p.325 ~ p.333
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Time-resolved Fluoroimmunoassay for the Measurement of 17beta-Estradiol using Anti-idiotypic Antibody
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Kim Yoon-Gyu
Lee Chi-Ho
Lee Won-Chang
Choi Young-Sook
Kim Jong-Bae
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Abstract
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A competitive type immunoassay method for 17beta-estradiol(E2) based on the idiotypic anti-idiotypic antibody and time-resolved fluorescence is described. The anti-idiotypic antibody(Ab2) produced to E2 binding site of the primary idiotype antibody (Ab1) was labelled with europium and was allowed to compete with E2 standards or serum sample for the binding sites of Ab1 which was bound to 2nd antibody captured ontothe surface of microtitre plates. Fluorescence measured by time-resolved fluorometer was inversely proportional to the concentration of E2 over the range 5~500pg/well. The sensitivity of the assay was 5pg per well which was compatible with that ofradioimmunoassay using the same Ab1 and 3H-E2 as a tracer. One great advantage of this method described here was to enable antibodies to be labelled instead of haptens, and thus makes it easier to develop sensitive and robust immunoassay systems specially for haptens
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