Terazawa K, Ji L Y, Mikami H, Togashi T, Takatori T
Department of Legal Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
J Immunoassay. 1991;12(3):413-24. doi: 10.1080/01971529108055080.
Monoclonal antibodies(moAbs) reactive with melatonin(MT) were produced using MT, coupled to bovine serum albumin(BSA) with the Mannich reaction, as immunogen and conventional hybridoma techniques. Hybridoma clones secreting the moAbs were selected by an enzyme-linked immunosorbent assay system using MT-carboxymethylchitin and BSA as screening antigens. The moAbs from 6 clones were characterized by a cross-reactivity test using radioimmunoassay with 125I-labelled MT. The moAbs recognized MT but hardly recognized other analogues except for N-acetylserotonin with a crossreactivity of 0.81%. An inhibition curve for MT was obtained in the range of 50 pg to 100 ng and 1.4 ng of MT inhibited the value of the assay by half. There is interference from some unknown source in human serum.
采用曼尼希反应将褪黑素（MT）与牛血清白蛋白（BSA）偶联作为免疫原，运用传统杂交瘤技术制备了与褪黑素（MT）反应的单克隆抗体（moAbs）。使用MT-羧甲基几丁质和BSA作为筛选抗原，通过酶联免疫吸附测定系统选择分泌moAbs的杂交瘤克隆。通过使用125I标记的MT进行放射免疫测定的交叉反应试验，对来自6个克隆的moAbs进行了表征。这些moAbs识别MT，但除了N-乙酰血清素外几乎不识别其他类似物，交叉反应率为0.81%。在50 pg至100 ng范围内获得了MT的抑制曲线，1.4 ng的MT可使测定值减半。人血清中存在来自某些未知来源的干扰。
