Concentrations of 2-hydroxyoestrogens in human sera measured by a heterologous immunoassay with an 125I-labelled ligand

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Abstract
A heterologous immunoassay for 2-hydroxyestrogens was established in which antibodies raised against 2-hydroxyestradiol-17-succinyl-bovine serum albumin serve as binding protein and 17-O-[N-.beta.-[125I]iodo-imidazolyl)ethylcarboxyamidomethyl]oximino-2,3-dihydroxy-1,3,5(10)-estratriene as radioligand. Lipophilic serum components competing for binding sites in this system were defined as total 2-hydroxyestrogens. The underlying assumption of specificity was supported by the pattern of cross-reactivity evaluated with structural related steroids and o-diphenols and byl the fact, that an additional chromatography of the serum extracts preceding the competing reaction had little if any effect. Sensitivity: 2.8 .+-. 1 pg/tube; accuracy: Y = 0.91x + 2.2; r = 0.989; precision: 5.8% intra-assay; 6.5% inter-assay. The following concentrations (.+-. standard deviation) were found in the sera of healthy subjects. Young men: 29 .+-. 5 pg/ml (n = 11); women follicular phase: 32 .+-. 8 pg/ml (n = 25); luteal phase: 53 .+-. 13 pg/ml (n = 23); postmenopausal women: 13 .+-. 4 pg/ml (n = 10); pregnant women 11th-20th week: 70 .+-. 16 mg/ml (n = 64); 36th-40th week: 240 .+-. 23 pg/ml (n = 40); newborn cord blood: 604 .+-. 43 pg/ml (n = 48).

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