Application of xanthine oxidase-catalyzed luminol chemiluminescence in a mouse interleukin-5 immunoassay

Autor: A. J. M. Van Oosterhout, H.A.H. Rongen, Auke Bult, H.M. van der Horst, W.P. van Bennekom
Rok vydání: 1996
Předmět:
Zdroj: Journal of Immunological Methods. 197:161-169
ISSN: 0022-1759
DOI: 10.1016/0022-1759(96)00131-7
Popis: 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.
Databáze: OpenAIRE