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用于测定紫杉醇及相关紫杉烷的超灵敏发光测定法的开发。

Development of a very sensitive luminescence assay for the measurement of paclitaxel and related taxanes.

作者信息

Guillemard V, Bicamumpaka C, Boucher N, Pagé M

机构信息

Department of Medical Biology, Faculty of Medicine, Laval University, Sainte-Foy, Quebec, Canada.

出版信息

Anticancer Res. 1999 Nov-Dec;19(6B):5127-30.

Abstract

A rapid and very sensitive enzyme immunoassay was developed for the measurement of paclitaxel and related taxanes in crude extracts of Taxus sp., in human serum and in culture medium of paclitaxel-producing microorganisms such as Erwinia taxi. For the ELISA, paclitaxel was chemically modified by the introduction of an amine to enable coupling with biotin. The presence of paclitaxel or related taxanes competitively inhibited the binding of paclitaxel-biotin to anti-taxane monoclonal antibody. This method detected paclitaxel in concentrations as low as 33 pM; the affinity of the antibody was higher for paclitaxel than for cephalomanine, baccatin and DAB. The sensitivity of this assay makes it useful for estimating the paclitaxel and taxanes content of Taxus sp. extracts, monitoring the paclitaxel serum level of paclitaxel treated patients and in other biological fluids.

摘要

开发了一种快速且非常灵敏的酶免疫测定法,用于测定红豆杉属植物粗提物、人血清以及产紫杉醇微生物(如紫杉欧文氏菌)培养基中的紫杉醇及相关紫杉烷。对于酶联免疫吸附测定(ELISA),通过引入胺对紫杉醇进行化学修饰,以实现与生物素的偶联。紫杉醇或相关紫杉烷的存在竞争性抑制了紫杉醇 - 生物素与抗紫杉烷单克隆抗体的结合。该方法能检测低至33皮摩尔浓度的紫杉醇;抗体对紫杉醇的亲和力高于对紫衫素、浆果赤霉素和二乙酰基巴卡亭的亲和力。该测定法的灵敏度使其可用于估算红豆杉属植物提取物中紫杉醇和紫杉烷的含量、监测紫杉醇治疗患者的血清紫杉醇水平以及其他生物体液中的含量。

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