Hacker M P, Dank J R, Ershler W B
Cancer Res. 1984 Feb;44(2):478-81.
Radioimmunoassays have been developed for pharmacokinetic studies of vinblastine. Although highly sensitive, radioimmunoassays are both expensive and potentially biohazardous. This paper describes a new immunoassay procedure, enzyme-linked immunosorbent assay, which uses enzymatic activity rather than radioactivity as an index of drug concentration. Antiserum to vinblastine was attached to the plastic wells of microtiter plates and incubated in the presence of varying amounts of vinblastine which had been conjugated to alkaline phosphatase. After brisk washing of the wells, p-nitrophenylphosphate was added to each well. The amount of enzyme present in the well was quantitated by the production of the chromophore, p-nitrophenol. The concentration of free vinblastine present in a given sample was inversely proportional to the enzymatic activity. The enzyme-linked immunosorbent assay is capable of detecting as little as 5 pg of either vinblastine or vincristine and is not cross-reactive with other commonly used oncolytic agents. A pharmacokinetic study was performed in rats administered vinblastine i.v., and a triphasic elimination curve was obtained. These results indicate that the enzyme-linked immunosorbent assay provides a nonradioactive, inexpensive, and sensitive method to monitor plasma levels of vinblastine. Further, since the antibody is cross-reactive with vincristine, it would appear that similar data could be generated for vincristine.
已开发出放射免疫分析法用于长春碱的药代动力学研究。放射免疫分析法虽然灵敏度高，但既昂贵又有潜在的生物危害性。本文描述了一种新的免疫分析方法，即酶联免疫吸附测定法，该方法使用酶活性而非放射性作为药物浓度指标。将抗长春碱的抗血清附着于微量滴定板的塑料孔中，并在存在与碱性磷酸酶偶联的不同量长春碱的情况下进行孵育。对孔进行快速洗涤后，向每个孔中加入对硝基苯磷酸酯。通过发色团对硝基苯酚的产生来定量孔中存在的酶量。给定样品中游离长春碱的浓度与酶活性成反比。酶联免疫吸附测定法能够检测低至5 pg的长春碱或长春新碱，并且与其他常用的溶瘤剂无交叉反应。对静脉注射长春碱的大鼠进行了药代动力学研究，并获得了三相消除曲线。这些结果表明，酶联免疫吸附测定法提供了一种非放射性、廉价且灵敏的方法来监测长春碱的血浆水平。此外，由于该抗体与长春新碱有交叉反应，因此似乎可以为长春新碱生成类似的数据。
