Choi J, Kim C, Choi M J
Doping Control Center, Korea Institute of Science and Technology, Seoul, South Korea.
J Chromatogr B Biomed Sci Appl. 1998 Feb 13;705(2):277-82. doi: 10.1016/s0378-4347(97)00527-6.
An accurate, simple and rapid immunoassay is demonstrated for the detection of methamphetamine in urine by capillary electrophoresis (CE) with laser-induced fluorescence (LIF). An aminobutyl derivative of methamphetamine was conjugated with proteins, and used as an immunogen to produce antibodies for the assay. The methamphetamine derivative was also labeled with fluorescein isothiocyanate (FITC) to compete with free methamphetamine in the sample for the antibody binding site. Levels of free and antibody-bound FITC-labeled methamphetamine were monitored by performing CE-LIF using an untreated fused-silica column. This competitive immunoassay used antiserum instead of purified antibody or antibody fragment, yet was found to have good precision with a sensitivity of lower than 20 ng/ml. Various antibodies were also screened, and cross-reactivity of anti-MA antibody with methamphetamine analogues were also investigated. The results indicate that CE-LIF-based immunoassay is a powerful tool for the screening and characterization of antibody and may have possible applications in the detection of abused drugs in urine.
展示了一种准确、简单且快速的免疫测定方法，用于通过毛细管电泳（CE）结合激光诱导荧光（LIF）检测尿液中的甲基苯丙胺。将甲基苯丙胺的氨基丁基衍生物与蛋白质偶联，并用作免疫原以产生用于该测定的抗体。甲基苯丙胺衍生物还用异硫氰酸荧光素（FITC）标记，以与样品中的游离甲基苯丙胺竞争抗体结合位点。使用未处理的熔融石英柱通过CE-LIF监测游离和抗体结合的FITC标记甲基苯丙胺的水平。这种竞争性免疫测定使用抗血清而非纯化抗体或抗体片段，但发现具有良好的精密度，灵敏度低于20 ng/ml。还筛选了各种抗体，并研究了抗甲基苯丙胺抗体与甲基苯丙胺类似物的交叉反应性。结果表明，基于CE-LIF的免疫测定是筛选和鉴定抗体的有力工具，并可能在尿液中滥用药物的检测中具有潜在应用。
