Development of an Enzyme Immunoassay for Serum 16-Dehydropregnenolone.
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- TAGAWA Noriko
- Clinical Chemistry Laboratory, Kobe Pharmaceutical University
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- SAIKI Kayoko
- Central Analytical Laboratory, Kobe Pharmaceutical University
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- KOBAYASHI Yoshiharu
- Clinical Chemistry Laboratory, Kobe Pharmaceutical University
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説明
We have developed an enzyme immunoassay (EIA) for serum 16-dehydropregnenolone (3β-hydroxy-5,16-pregnadien-20-one; 16-DHP). The antiserum against 16-DHP-3-hemisuccinate conjugated bovine serum albumin (16-DHP-3HS-BSA) was raised in rabbits. For use as an enzyme labeled antigen, 16-DHP-3HS was conjugated to alkaline phosphatase. The minimal amount of 16-DHP detected was 4pg (0.013 pmol)/assay and the measurable range was from 0.06—60 ng/ml (0.191—191 nmol/l). The intra-assay coefficient of variation (C.V.) was 4.1% (0.73±0.03 ng/ml, mean±S.D., n=6), and inter-assay C.V. was 7.7% (0.13±0.01 ng/ml, n=6). A liner relation was observed between the serum sample dilution and the 16-DHP concentration. For the recovery study, authentic 16-DHP was added to a serum sample (original concentration: 0.10—0.14 ng/ml), and the recovery was found to be 94.4—96.8% (final 16-DHP concentrations calculated: 0.29—16.3 ng/ml). To investigate the reliability of the present EIA, the values from our EIA were compared with those obtained by GC-MS. The 16-DHP concentration could not be measured in serum by GC-MS because of its sensitivity. Therefore, the conjugated steroid, 16-DHPS, was first enzymatically hydrolysed and then the 16-DHP measured by both methods. There was a good correlation between the levels determined by these methods (Pearson’s correlation coefficient: r=0.927, p<0.001, y=0.74x+3.61, n=27). The serum concentrations of 16-DHP in neonates and umbilical vein were 0.53±0.09 ng/ml and 0.88±0.61 ng/ml, respectively. No 16-DHP was detected in serum from normal healthy adults using the present EIA. These results suggest that 16-DHP originates from the fetus and neonate.
収録刊行物
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- Biological & Pharmaceutical Bulletin
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Biological & Pharmaceutical Bulletin 24 (8), 867-871, 2001
公益社団法人 日本薬学会
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詳細情報 詳細情報について
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- CRID
- 1390001204625184640
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- NII論文ID
- 110003638578
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- NII書誌ID
- AA10885497
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- COI
- 1:CAS:528:DC%2BD3MXlvVCms7k%3D
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- ISSN
- 13475215
- 09186158
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- NDL書誌ID
- 5852067
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- PubMed
- 11510475
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- 使用不可