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一种用于检测抗肿瘤聚乙炔醇——人参三醇的高灵敏度酶联免疫吸附测定(ELISA)。

A highly sensitive enzyme-linked immunosorbent assay (ELISA) for antitumor polyacetylenic alcohol, panaxytriol.

作者信息

Saita T, Matsunaga H, Yamamoto H, Nagumo F, Fujito H, Mori M, Katano M

机构信息

Faculty of Hospital Pharmacy, Saga, Japan.

出版信息

Biol Pharm Bull. 1994 Jun;17(6):798-802. doi: 10.1248/bpb.17.798.

Abstract

A new type of antitumor polyacetylenic alcohol, panaxytriol, was isolated from the roots of Panax ginseng C. A. Meyer. A highly sensitive enzyme-linked immunosorbent assay (ELISA) for the determination of panaxytriol was developed, which is capable of measuring as low as 25.6 pg/ml. Anti-panaxytriol antibody was obtained by immunizing rabbits with panaxytriol conjugated with bovine serum albumin using the N-succinimidyl ester method. An enzyme marker was similarly prepared by coupling panaxytriol with horseradish peroxidase. The specificity of this ELISA seems to be primarily toward both the glycol moiety and the diacetylene moiety of the panaxytriol, showing a slight cross-reaction with the other panaxytriol analogues which are structurally different only in C-9,10 positions, but no cross-reaction with the 1,2-decanediol or 3-nonyn-1-ol. The values for panaxytriol concentration detected by this assay were comparable with those detected by the gas chromatography method. The ELISA was about 5000 times more sensitive in detecting panaxytriol. Using this assay, panaxytriol levels were easily determined in the blood of rats. The ELISA may be a valuable tool for studies of the biological and pharmacological properties of the polyacetylenic alcohol, panaxytriol.

摘要

从人参(Panax ginseng C. A. Meyer)根中分离出一种新型抗肿瘤聚乙炔醇——人参三醇。开发了一种用于测定人参三醇的高灵敏度酶联免疫吸附测定法(ELISA),该方法能够检测低至25.6 pg/ml的人参三醇。通过使用N-琥珀酰亚胺酯法将与人血清白蛋白偶联的人参三醇免疫兔子,获得了抗人参三醇抗体。同样通过将人参三醇与辣根过氧化物酶偶联制备了酶标记物。这种ELISA的特异性似乎主要针对人参三醇的二醇部分和二乙炔部分,与仅在C-9、10位结构不同的其他人参三醇类似物有轻微交叉反应,但与1,2-癸二醇或3-壬炔-1-醇无交叉反应。该测定法检测到的人参三醇浓度值与气相色谱法检测到的值相当。ELISA检测人参三醇的灵敏度高出约5000倍。使用该测定法,可以轻松测定大鼠血液中的人参三醇水平。ELISA可能是研究聚乙炔醇人参三醇的生物学和药理学特性的有价值工具。

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