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一种用于测定红豆杉属植物组织和人体血浆中紫杉醇及紫杉烷的酶免疫测定法。

An enzyme immunoassay for the determination of taxol and taxanes in Taxus sp. tissues and human plasma.

作者信息

Grothaus P G, Raybould T J, Bignami G S, Lazo C B, Byrnes J B

机构信息

Hawaii Biotechnology Group, Inc., Aiea 96701.

出版信息

J Immunol Methods. 1993 Jan 14;158(1):5-15. doi: 10.1016/0022-1759(93)90253-4.

Abstract

A rapid and sensitive indirect competitive inhibition enzyme immunoassay (CIEIA) method has been developed for quantitating taxanes in extracts of Taxus brevifolia Nutt. tissue and in human plasma. High titer rabbit polyclonal antibodies (pAb) were raised against a conjugate of 7-succinyltaxol and keyhole limpet hemocyanin. The presence of taxane analyte competitively inhibited the binding of the rabbit anti-taxane pAbs to a 7-succinyltaxol-bovine serum albumin solid phase coating antigen. The CIEIA detected taxol and cephalomannine in concentrations as low as 0.3 ng/ml (3.5 x 10(-4) microM), but did not detect baccatin III or 10-deacetylbaccatin III at concentrations below 1000 ng/ml (1.7 microM and 1.8 microM, respectively). This method is useful for estimating the taxane content of T. brevifolia extracts and may be useful for monitoring the taxol plasma level of taxol-treated patients.

摘要

已开发出一种快速灵敏的间接竞争抑制酶免疫分析(CIEIA)方法,用于定量短叶红豆杉组织提取物和人血浆中的紫杉烷。用7-琥珀酰紫杉醇与钥孔戚血蓝蛋白的偶联物制备了高效价兔多克隆抗体(pAb)。紫杉烷分析物的存在竞争性抑制兔抗紫杉烷pAb与7-琥珀酰紫杉醇-牛血清白蛋白固相包被抗原的结合。CIEIA检测紫杉醇和三尖杉宁碱的浓度低至0.3 ng/ml(3.5×10⁻⁴ μM),但在浓度低于1000 ng/ml(分别为1.7 μM和1.8 μM)时未检测到巴卡亭III或10-去乙酰巴卡亭III。该方法可用于估计短叶红豆杉提取物中的紫杉烷含量,也可能有助于监测紫杉醇治疗患者的血浆紫杉醇水平。

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