Rongen H A, van der Horst H M, van Oosterhout A J, Bult A, van Bennekom W P
Department of Pharmaceutical Analysis, Faculty of Pharmacy, Utrecht University, Netherlands.
J Immunol Methods. 1996 Oct 16;197(1-2):161-9. doi: 10.1016/0022-1759(96)00131-7.
A chemiluminescent substrate reagent for use in a sandwich immunoassay for the model antigen mouse interleukin-5 (IL-5) was developed using xanthine oxidase and luminol. Various parameters involved in this chemiluminescent reaction have been studied, including the substrate hypoxanthine, luminol and the Fe(II)-EDTA complex. Addition of the Fe(II)-EDTA complex enhances the chemiluminescence signal considerably. The xanthine oxidase-catalyzed chemiluminescent immunoassay was compared to horseradish peroxidase-linked immunoassays with luminol as chemiluminescent, and tetramethyl benzidine as colorimetric substrate. The detection limit of the xanthine oxidase-luminol assay was found to be about 0.6 pg/ml IL-5, whereas the peroxidase-catalyzed immunoassays have detection limits of about 1.3 (HRP-TMB) and 2.9 pg/ml (HRP-luminol) IL-5.
使用黄嘌呤氧化酶和鲁米诺开发了一种用于模型抗原小鼠白细胞介素-5（IL-5）夹心免疫测定的化学发光底物试剂。已经研究了该化学发光反应中涉及的各种参数，包括底物次黄嘌呤、鲁米诺和Fe（II）-EDTA络合物。添加Fe（II）-EDTA络合物可显著增强化学发光信号。将黄嘌呤氧化酶催化的化学发光免疫测定与以鲁米诺为化学发光底物、四甲基联苯胺为比色底物的辣根过氧化物酶连接免疫测定进行了比较。发现黄嘌呤氧化酶-鲁米诺测定的检测限约为0.6 pg/ml IL-5，而过氧化物酶催化的免疫测定的检测限约为1.3（HRP-TMB）和2.9 pg/ml（HRP-鲁米诺）IL-5。
