人表皮生长因子的超灵敏时间分辨免疫荧光分析

An ultrasensitive time-resolved immunofluorometric assay of human epidermal growth factor.

作者信息

Pesonen K, Alfthan H, Stenman U H, Viinikka L, Perheentupa J

出版信息

Anal Biochem. 1986 Sep;157(2):208-11. doi: 10.1016/0003-2697(86)90617-2.

Abstract

We have developed a sandwich-type time-resolved immunofluorometric assay (TR-IFMA) for human epidermal growth factor (hEGF) in body fluids. A two-step solid-phase technique was used. The assay utilizes a polyclonal anti-hEGF attached to the solid phase, and a monoclonal anti-hEGF labeled with Europium (III) as a tracer. The sensitivity of the assay (2.5 pg/ml) is at least 20 times better than what has been achieved by radioimmunoassay (RIA), and the measuring range is much wider: 2.5-5000 pg/ml. The feasibility of TR-IFMA was tested by assaying urine containing large amounts and amniotic fluid containing small amounts (mostly undetectable by RIA) of immunoreactive hEGF. The correlation between urine hEGF concentrations (1-100 ng/ml) measured by RIA and TR-IFMA was good: r = 0.96.

摘要

我们开发了一种用于检测体液中人表皮生长因子（hEGF）的夹心型时间分辨免疫荧光分析（TR-IFMA）方法。采用了两步固相技术。该分析方法利用固定在固相上的多克隆抗hEGF抗体，以及用铕（III）标记的单克隆抗hEGF抗体作为示踪剂。该分析方法的灵敏度（2.5 pg/ml）比放射免疫分析（RIA）至少高20倍，测量范围也更宽：2.5 - 5000 pg/ml。通过检测含有大量免疫反应性hEGF的尿液和含有少量（大多RIA检测不到）免疫反应性hEGF的羊水，测试了TR-IFMA的可行性。RIA和TR-IFMA测量的尿hEGF浓度（1 - 100 ng/ml）之间的相关性良好：r = 0.96。