大鼠腺体激肽释放酶的过量抗体免疫测定法。测定与抑制剂结合的激肽释放酶及血浆中的激肽释放酶。

Excess antibody immunoassay for rat glandular kallikrein. Measurement of kallikrein complexed with inhibitors and in plasma.

作者信息

Johansen L, Nustad K, Berg T, Pierce J V

出版信息

J Immunol Methods. 1984 Apr 27;69(2):253-66. doi: 10.1016/0022-1759(84)90323-5.

PMID:6201559

Abstract

We have recently developed an immunoradiometric assay (IRMA) for specific measurement of immunoreactive kallikrein which allows a simultaneous determination of the enzymatic activity of kallikrein. This paper describes the application of this method for measurements of glandular kallikrein complexed with inhibitors. Interference by low molecular weight inhibitors such as benzamidine and Trasylol was easily overcome by increasing the amount of immobilized anti-kallikrein antibody added in the assay, and by prolonging the incubation time of the antigen-binding step. The recovery of kallikrein in complex with plasma inhibitors was complete only when the anti-kallikrein antibody was immunoadsorbed onto a solid-phase sheep anti-rabbit immunoglobulin. The dose-response curve of glandular kallikrein in plasma paralleled that of purified kallikrein in both the immunoradiometric and the immunoenzymometric assays. The concentration of immunoreactive glandular kallikrein in normal rat plasma was 12.8 +/- 4.3 nU/ml. The enzymatic activity of this immunoreactive kallikrein was 86% inhibited.

摘要

我们最近开发了一种免疫放射分析方法（IRMA），用于特异性测量免疫反应性激肽释放酶，该方法能够同时测定激肽释放酶的酶活性。本文描述了该方法在测量与抑制剂复合的腺体激肽释放酶中的应用。通过增加测定中加入的固定化抗激肽释放酶抗体的量，以及延长抗原结合步骤的孵育时间，很容易克服诸如苯甲脒和抑肽酶等低分子量抑制剂的干扰。只有当抗激肽释放酶抗体免疫吸附到固相羊抗兔免疫球蛋白上时，与血浆抑制剂复合的激肽释放酶的回收率才是完全的。在免疫放射分析和免疫酶分析中，血浆中腺体激肽释放酶的剂量反应曲线与纯化激肽释放酶的曲线平行。正常大鼠血浆中免疫反应性腺体激肽释放酶的浓度为12.8±4.3 nU/ml。这种免疫反应性激肽释放酶的酶活性被抑制了86%。