Fluorescence polarization immunoassay of progesterone

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Title
Fluorescence polarization immunoassay of progesterone
Author(s)
Myung Ja Choi; Jeong Eun Choi; Do Young Yoon; Jong Sei Park; Sergei A Eremin
Bibliographic Citation
Biological & Pharmaceutical Bulletin, vol. 20, no. 4, pp. 309-314
Publication Year
1997
Abstract
A homogeneous fluorescence polarization immunoassay (FPIA) was developed to measure levels of progesterone in urine using a TDx analyzer in photocheck mode (Abbott Labs). Two tracers of ethylenediamine fluorescein thiocarbamyl (EDF) were employed; one was synthesized from 11α-hydroxyhemisuccinate progesterone (Prog11OH-HS) and the other was synthesized from 3-(o- carboxymethyl)oxime progesterone (Prog-3CMO). Each derivative of progesterone was conjugated with bovine serum albumin and used as an immunogen which produced monoclonal antibody clone 15A (MAb 15A, anti-Prog-11OH-HS) and clone 2B7 (MAb 2B7, anti-Prog-3CMO), respectively. Different combinations of tracers and antibodies were investigated in the FPIA system. Similar sensitivity was observed when using the pair, MAb 2B7 and its homologous tracer, Prog-3CMO-EDF, or MAb 15A and its homologous tracer, Prog-11OH-HS- EDF. In this immunoassay, no separation step was required and the total time for an assay of 10 samples was approximately 7 min. The progesterone detection limit in a 10 μl sample was 3 ng/ml. The cross-reactivity results indicate that the A-, B- and D-ring of asteroid are buried in the binding pocket of MAb 15A, while the C-ring faced outward, resulting in cross- reactivity with 11-αhydroxy progesterone. The A-, B- and C-ring of asteroid of MAb 2B7, in contrast, are buried deep in the pocket leaving the D-ring facing outward, resulting in some different degrees of cross-reactivity with C17 position substituted steroids.
Keyword
fluorescence polarization immunoassayprogesteronefluorescenceimmunoassaytracerimmunogen
ISSN
0918-6158
Publisher
Pharmaceutical Soc Japan
DOI
http://dx.doi.org/10.1248/bpb.20.309
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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