一种用于测量神经降压素的高灵敏度和高选择性放射免疫分析方法。

A highly sensitive and selective radioimmunoassay for the measurement of neurotensin.

作者信息

Holtom P E, Wasuntarawat C, Moss S H, Aspley S, Needham P L, Bennett G W

机构信息

School of Biomedical Sciences (E floor), Queen's Medical Centre, NG7 2UH, Nottingham, UK.

出版信息

J Neurosci Methods. 2000 Jul 31;100(1-2):151-6. doi: 10.1016/s0165-0270(00)00243-0.

DOI:10.1016/s0165-0270(00)00243-0

PMID:11040378

Abstract

A highly selective and sensitive radioimmunoassay (RIA) for the detection of endogenous neurotensin (NT) has been developed. We have raised a C-terminally-directed antibody (CAb) that specifically binds 'biologically active' NT (NT and NT(8-13)) and that does not significantly cross-react with inactive NT metabolites or other bioactive peptides in the CNS. By reducing the volume of the assay to a low volume-RIA (30 microl), such that in vivo measurements can be made, we have increased the sensitivity (<0.3 fmol per tube), with inter- and intra-assay variations of 11.2 and 5.8%, respectively. Comparisons with similar methods of detecting NT have demonstrated that this RIA has a higher sensitivity than previously used RIA's and ELISA's. The data presented suggests that this sensitive RIA is a reliable method ideal for the detection of small quantities of biologically active NT.

摘要

已开发出一种用于检测内源性神经降压素（NT）的高选择性和高灵敏度放射免疫分析法（RIA）。我们制备了一种C端导向抗体（CAb），它能特异性结合“生物活性”NT（NT和NT(8 - 13)），且与中枢神经系统中无活性的NT代谢产物或其他生物活性肽无明显交叉反应。通过将检测体积减小至低体积RIA（30微升），从而能够进行体内测量，我们提高了灵敏度（每管<0.3飞摩尔），批间和批内变异分别为11.2%和5.8%。与检测NT的类似方法比较表明，这种RIA比以前使用的RIA和酶联免疫吸附测定（ELISA）具有更高的灵敏度。所呈现的数据表明，这种灵敏的RIA是检测少量生物活性NT的理想可靠方法。