Yin B Y, Gasparro F P, Bevilacqua P M, Santella R M
Division of Environmental Sciences, School of Public Health, Columbia University, New York, NY 10032.
J Invest Dermatol. 1991 Dec;97(6):1001-4. doi: 10.1111/1523-1747.ep12492190.
An immunologic method for the quantitation of 8-methoxypsoralen (8-MOP) levels in human plasma has been developed. A monoclonal antibody recognizing 8-MOP was prepared by immunizing mice with an 8-MOP derivative conjugated to bovine serum albumin with 1-ethyl-3-(3-dimethylamino-propyl)-carbodiimide-HCl. The antibody was characterized by competitive enzyme-linked immunosorbent assay (ELISA) and recognizes 8-MOP (50% inhibition at 2 pmol) as well as structurally related psoralen derivatives including 4'-aminomethyl-4,5',8-trimethylpsoralen (50% inhibition at 50 pmol), 5-methoxypsoralen (50% inhibition at 150 pmole), and 6,4,4'-trimethylangelicin (50% inhibition at 360 pmol). The assay has a limit of sensitivity of 1 ng/ml plasma. For analysis of 8-MOP levels in plasma, samples were first extracted using SepPak C18 cartridges. The extracts were analyzed for 8-MOP levels both by ELISA and high-pressure liquid chromatography. There was a good correlation between the values determined by both methods (r = 0.92, p less than 0.0005). The development of immunologic methods should greatly facilitate the quantitation of 8-MOP levels in patient plasma.
已开发出一种用于定量人血浆中8-甲氧基补骨脂素（8-MOP）水平的免疫方法。通过用与牛血清白蛋白偶联的8-MOP衍生物（用1-乙基-3-（3-二甲基氨基丙基）-碳二亚胺盐酸盐）免疫小鼠，制备了识别8-MOP的单克隆抗体。该抗体通过竞争性酶联免疫吸附测定（ELISA）进行表征，可识别8-MOP（在2 pmol时50%抑制）以及结构相关的补骨脂素衍生物，包括4'-氨基甲基-4,5',8-三甲基补骨脂素（在50 pmol时50%抑制）、5-甲氧基补骨脂素（在150 pmol时50%抑制）和6,4,4'-三甲基当归素（在360 pmol时50%抑制）。该测定法的血浆灵敏度极限为1 ng/ml。为了分析血浆中的8-MOP水平，首先使用SepPak C18柱对样品进行提取。通过ELISA和高压液相色谱法对提取物中的8-MOP水平进行分析。两种方法测定的值之间具有良好的相关性（r = 0.92，p < 0.0005）。免疫方法的开发应极大地促进患者血浆中8-MOP水平的定量。
