<原著>Sex-steroid-binding plasma protein(SBP)の基礎的および臨床的研究 : (1) : SBPのradioimmunoassay(RIA)の開発とSBPの安定性に関する基礎的研究
スポンサーリンク
概要
- 論文の詳細を見る
Human sex-steroid-binding plasma protein (SBP) is a protein to bind testosterone, estradiol and dihydrotestosterone. In human plasma, SBP regulates free sex steroids which are biologically active. The purpose of this study is to develop a radioimmunoassay (RIA) of SBP for clinical use and thereby to examine the stability of SBP under various conditions including its changes in acidity and temperature. Human SBP was isolated from pooled serum with affinity chromatography and subsequently with polyacrylamide get electrophoresis. These procedures yielded a purified SBP preparation, concentrated 6,000 fold over original pooled serum with an overall recovery of 7%. Purified SBP showed a single band in polyacrylamide gel electrophoresis. Specific antibody against human SBP was raised in a rabbit by using the obtained SBP preparation, and its specificity to SBP was demonstrated by immunoelectrophoresis. Get filtration pattern of ^3H-dihydrotestosterone-SBP=anti human SBP antibody revealed that anti-human SBP antibody bound to ^3H-DHT-bound SBP and not to ^3H-DHT-bound albumin. A RIA for SBP was constructed based on a double antibody method by utilizing anti rabbit Ig G in a goat as a second antibody. The intraassay coefficient of variation (CV%) for a normal male and a normal female was 2.1 and 2.5 and the intraassay coefficient of variation for them was 7.0 and 5.3,respectively. A dilution test of our RIA demonstrated a good recovery. In the present anti-SBP antibody, no cross reactivity was observed with any of transferrin, albumin, testosterone, estradiol and dihydrotestosterone. The SBP levels determined by the present RIA showed a good correlation with those determined by dextran-coated charcoal assay. The stability of SBP in various conditions was studied with both the present RIA and the binding assay above. SBP values, measured by both methods were stable between pH 6-9 for two weeks, but decreased in parallel at the pH of the pHs both 11 or more and 4 and downward in 24 hours. With respect to the effects of temperature, SBP values measured with both methods were stable for six months at -20℃, and were also stable at room temperature and 37℃ for two weeks. However, SBP values measured by both methods decayed in parallel within 1 hour at 56℃. Preliminary results of SBP measurement by our RIA were 1.3±0.1 (μg/ml)(mean±SE) in twenty normal males and 3.3±0.8 (μg/ml) in twenty-seven normal females, respectively.
- 近畿大学の論文
- 1985-12-25
著者
関連論文
- 32.糖尿病におけるHbA_1測定の意義
- 糖代謝異常を来す諸疾患における尿中C-peptide量
- 34.下垂体性, 副腎腺腫性, および両側性結節性副腎過形成性, 各クッシング症候群の予後に関する比較検討
- Sex-steroid-binding plasma protein(SBP)の基礎的および臨床的研究 : (2) : 健常者および肝硬変症者におけるSBPおよび各種性腺ホルモンの測定
- Sex-steroid-binding plasma protein(SBP)の基礎的および臨床的研究 : (1) : SBPのradioimmunoassay(RIA)の開発とSBPの安定性に関する基礎的研究
- 4端子Bipolar凝固子による経内視鏡的高周波電気凝固止血法の基礎的検討