Duncan M E, McAleese S M, Booth N A, Melvin W T, Fothergill J E
Department of Molecular and Cell Biology, University of Aberdeen, UK.
J Immunol Methods. 1992 Jul 6;151(1-2):227-36. doi: 10.1016/0022-1759(92)90121-9.
Monoclonal antibodies specific for the gamma isozyme of human enolase (known as neuron-specific enolase or NSE) have been raised against synthetic peptides after coupling to carrier protein: the selected peptides were those corresponding to regions of amino acid sequence difference between the alpha and gamma subunits of these closely similar isozymes. This technique gave monoclonal antibodies of high specificity and affinity. Two monoclonal antibodies raised against different peptides were used to develop a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA), using one as the solid-phase antibody and the other conjugated to horseradish peroxidase to detect the bound NSE. This assay provides a simple and routine method of detecting NSE in serum samples from patients with small-cell carcinoma of the lung and related tumours.
已针对与载体蛋白偶联后的合成肽产生了对人烯醇化酶γ同工酶(称为神经元特异性烯醇化酶或NSE)具有特异性的单克隆抗体:所选肽对应于这些极为相似的同工酶的α和γ亚基之间氨基酸序列差异的区域。该技术产生了具有高特异性和亲和力的单克隆抗体。使用针对不同肽产生的两种单克隆抗体开发了一种双抗体夹心酶联免疫吸附测定法(ELISA),其中一种用作固相抗体,另一种与辣根过氧化物酶偶联以检测结合的NSE。该测定法提供了一种简单且常规的方法,用于检测来自肺癌和相关肿瘤患者血清样本中的NSE。