Fluorescence Polarization Immunoassay of Progesterone
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概要
- 論文の詳細を見る
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 (Prog-11OH-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 approximaterly 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 a steroid 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 a steroid 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.
- 公益社団法人日本薬学会の論文
- 1997-04-15
著者
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Choi M
Bioanalysis & Biotransformation Research Center Korea Institute Of Science & Technology
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Choi Myung
Doping Control Center Korea Institute Of Science And Technology
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PARK Jongsei
Doping Control Center, Korea Institute of Science and Technology
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Yoon Do
Bioscience & Biotechnology Research Institute Korea Institute Of Science And Technology
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Eremin Sergei
Doping Control Center Korea Institute Of Science And Technology:(present Address)department Of Chemi
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Park Jisoo
R & D Center Daewoong Pharmaceutical Co. Ltd.
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Park Jongsei
Doping Control Center Korea Institute Of Science And Technology
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Choi Jeongeun
Doping Control Center Korea Institute Of Science And Technology
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Choi J
Doping Control Center Korea Institute Of Science And Technology
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