Terazawa K, Ji L Y, Takatori T, Aoki K, Hirose Y, Kuroiwa Y
Department of Legal Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
J Immunoassay. 1991;12(2):277-92. doi: 10.1080/01971529108055072.
Monoclonal antibodies (McAbs) specific to methamphetamine (MA) were produced using p-amino MA coupled to bovine serum albumin (BSA) with glutaraldehyde (GA) as an immunogen and with conventional hybridoma techniques. Hybridoma clones secreting the McAbs were selected by an enzyme-linked immunosorbent assay (ELISA) system using both the above conjugate and BSA modified with GA as screening antigens. In the ELISA system were used avidin and biotinyl-alkaline phosphatase which converts nicotinamide adenine dinucleotide phosphate (NADP) into NAD. The final enzyme activity was determined using diformazan of nitroblue tetrazolium formed together with the NAD produced, alcohol dehydrogenase and phenazine methosulfate. The McAbs from 9 clones were characterized by a crossreactivity test using the ELISA. The McAbs recognized MA (100%), methoxyphenamine (8.0%), ephedrine (2.3%), but did not react with metylephedrine, amphetamine, OH-amphetamine, dimethylamphetamine, beta-phenylethylamine, norephedrine, phentermine and ranitidine. An inhibition curve for MA was obtained in the range of 0.75 to 50 ng.
采用对氨基甲基苯丙胺与牛血清白蛋白（BSA）通过戊二醛（GA）偶联作为免疫原，运用传统杂交瘤技术制备了对甲基苯丙胺（MA）具有特异性的单克隆抗体（McAbs）。使用上述偶联物和经GA修饰的BSA作为筛选抗原，通过酶联免疫吸附测定（ELISA）系统筛选分泌McAbs的杂交瘤克隆。在ELISA系统中使用了抗生物素蛋白和生物素化碱性磷酸酶，该酶可将烟酰胺腺嘌呤二核苷酸磷酸（NADP）转化为NAD。最终的酶活性通过与产生的NAD一起形成的硝基蓝四氮唑的双甲臜、乙醇脱氢酶和硫酸甲基吩嗪来测定。使用ELISA通过交叉反应试验对来自9个克隆的McAbs进行了表征。这些McAbs识别MA（100%）、甲氧苯丙胺（8.0%）、麻黄碱（2.3%），但与甲基麻黄碱、苯丙胺、羟基苯丙胺、二甲基苯丙胺、β-苯乙胺、去甲麻黄碱、苯丁胺和雷尼替丁不发生反应。在0.75至50 ng范围内获得了MA的抑制曲线。
