采用单克隆抗IgE抗体的斑点免疫结合试验检测和定量人IgE。

Dot-immunobinding assay with monoclonal anti-IgE antibodies for the detection and quantitation of human IgE.

作者信息

Hong C S, Stadler B M, Wälti M, De Weck A L

出版信息

J Immunol Methods. 1986 Dec 24;95(2):195-202. doi: 10.1016/0022-1759(86)90406-0.

DOI:10.1016/0022-1759(86)90406-0

Abstract

This paper describes a dot immunobinding assay for determining total human IgE with a tandem of monoclonal anti-IgE antibodies. Minute quantities of monoclonal anti-IgE antibodies were adsorbed on nitrocellulose discs. IgE bound to this solid phase monoclonal anti-IgE antibody was detected by a second monoclonal antibody conjugated with horseradish peroxidase. Using 4-chloro-1-naphthol as a chromogen results in a stable colour reaction that can be semiquantitatively analysed by the naked eye. The colour intensities of the reaction were also analysed by densitometry, yielding a very reproducible quantitation of human serum IgE. Using a serum dilution of 1:50, IgE could be detected in the range of 12.5-2500 U/ml. Using non-diluted serum samples IgE levels between 0.05-50 U/ml were reproducibly measured. Total serum IgE as determined by this dot assay correlated very well with IgE determinations performed by the commercial PRIST assay.

摘要

本文描述了一种采用串联单克隆抗IgE抗体测定人总IgE的斑点免疫结合测定法。将微量单克隆抗IgE抗体吸附在硝酸纤维素膜上。通过与辣根过氧化物酶偶联的第二种单克隆抗体检测结合到该固相单克隆抗IgE抗体上的IgE。使用4-氯-1-萘酚作为显色剂可产生稳定的显色反应，该反应可用肉眼进行半定量分析。还通过光密度测定法分析反应的颜色强度，从而对人血清IgE进行非常可重复的定量。使用1:50的血清稀释度，可在12.5 - 2500 U/ml范围内检测到IgE。使用未稀释的血清样本，可重复测量0.05 - 50 U/ml之间的IgE水平。通过这种斑点测定法测定的总血清IgE与通过商业PRIST测定法进行的IgE测定结果相关性非常好。