Solid phase fluoroimmunoassay for 11-deoxycortisol in serum using 21-amino-17-hydroxyprogesterone.
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.
D015065:17-Hydroxycorticosteroids / Q000097:blood*; D003350:Cortodoxone / Q000097:blood*; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006801:Humans; D006908:Hydroxyprogesterones / Q000138:chemical synthesis*; D007118:Immunoassay / Q000379:methods; D007202:Indicators and Reagents; D008797:Metyrapone; D011863:Radioimmunoassay / Q000379:methods; D013861:Thiocyanates
D015065:17-Hydroxycorticosteroids; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006908:Hydroxyprogesterones; D007202:Indicators and Reagents; D013861:Thiocyanates; C037605:21-amino-17-hydroxyprogesterone; D016650:Fluorescein-5-isothiocyanate; D003350:Cortodoxone; D008797:Metyrapone