Solid phase fluoroimmunoassay for 11-deoxycortisol in serum using 21-amino-17-hydroxyprogesterone |
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Authors: | F Watanabe N Tsubota Y Kobayashi O Miyata I Ninomiya K Miyai |
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Institution: | 1. Clinical Chemistry Laboratory 4-19-1, Motoyamakitamachi, Higashinada-ku, Kobe, Japan;2. Medicinal Chemistry Laboratory of Kobe Women''s College of Pharmacy, 4-19-1, Motoyamakitamachi, Higashinada-ku, Kobe, Japan;3. Central Laboratory for Clinical Investigation and Department of Laboratory Medicine, Osaka University Medical School, 1-1-50, Fukushima, Fukushima-ku, Osaka, Japan |
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Abstract: | Solid phase fluoroimmunoassay of serum 11-deoxycortisol (17,21-dihydroxy-4-pregnene-3,20-dione) was established using fluorescein isothiocyanate-labelled 11-deoxycortisol and anti-11-deoxycortisol antibody-conjugated polyacrylamide beads. 21-Amino-17-hydroxyprogesterone (21-amino-17-hydroxy-4-pregnene-3,20-dione) was synthesized as a useful derivative for preparing the fluorescent dye conjugate. Serum 11-deoxycortisol was measured with this assay system after extraction and purification by Sephadex LH-20 column chromatography. The minimal amount of 11-deoxycortisol detected was 40 pg/tube and the measurable range was from 0.04 to 5.0 microgram/dl. Intra- and inter-assay coefficients of variation were 8.3% (n=6) and 9.8% (n=5), respectively. 11-deoxycortisol values determined by the present assay correlated well with those determined by radioimmunoassay. The present assay is particularly suitable for estimating the conditions of the pituitary and adrenocortical functions. |
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