A radioimmunoassay for serum dehydroepiandrosterone sulfate
A radioimmunoassay for serum dehydroepiandrosterone sulfate (DHEAS) (1) has been developed using anti-DHEA antiserum obtained by immunizing rabbits with DHEA-17 oxime-bovine serum albumin. Serum volume of 0.01 to 0. 1 ml was used for analysis. After the addition of ammonium salt of DHEA-7α 3 H sulfa...
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Published in | Steroids Vol. 20; no. 6; pp. 813 - 824 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
United States
Elsevier Inc
01.12.1972
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Subjects | |
Online Access | Get full text |
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Summary: | A radioimmunoassay for serum dehydroepiandrosterone sulfate (DHEAS) (1) has been developed using anti-DHEA antiserum obtained by immunizing rabbits with DHEA-17 oxime-bovine serum albumin. Serum volume of 0.01 to 0. 1 ml was used for analysis. After the addition of ammonium salt of DHEA-7α
3 H sulfate for recovery and a preliminary removal of DHEA, DHEAS was extracted as pyridinium salt by methylene chloride. The dried extract was subjected to solvolysis (Burstein & Lieberman), followed by paper chromatography. The eluates and DHEA-7 α
3H which was added to determine the % free of DHEA were evaporated and incubated with the antiserum containing pepsin treated human immune serum globulin and bovine serum albumin at 37°C for 1 hour. Ammonium sulfate was used to separate free from bound DHEA. The accuracy, precision and specificity were satisfactory. The sensitivity was 3 ng per sample. The blank values could not be differentiated from zero. Although the antiserum reacts with the other 3βOHΔ
5 steroids as well as DHEA, the complete separation of DHEA from the other 3βOHΔ
5 steroids was achieved chromatographically. Serum DHEAS levels in normal subjects and patients with adrenocortical disorders obtained with the radioimmunoassay were comparable to those obtained with gasliquid chromatography. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0039-128X 1878-5867 |
DOI: | 10.1016/0039-128X(72)90059-1 |