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使用单分子阵列超敏感检测酶活性

Xu Wang^1,2, Alana F Ogata^1,2, David R Walt^1,2

¹Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, United States.

Journal of the American Chemical Society

|August 17, 2020

概括

使用单分子阵列 (eSimoa) 的新型超敏感酶测定为酶活性提供了前所未有的检测灵敏度. 这种方法促进了酶研究,药物发现和诊断.

科学领域:

生物化学
分子生物学
测试开发

背景情况:

酶测试对于诊断,蛋白质组学和药物发现至关重要.
现有的方法往往缺乏足够的分析灵敏度.

研究的目的:

开发一种超敏感的方法来检测酶活性.
证明该方法对各种酶和抑制剂选的适用性.

主要方法:

使用单分子阵列 (eSimoa) 来检测酶活性.
用于单个分子分析,将合基质转化为磁性珠.
对蛋白激酶,端粒酶,SET7/9和GalNAc-T进行了测试.

主要成果:

在检测酶活性方面取得了前所未有的灵敏度.
成功确定了博苏替尼和达沙替尼的抑制常数.
在单细胞水平上启用激酶活性测量.

结论:

该eSimoa测试是一种简单,快速和高度敏感的酶检测平台.
该方法可扩展到广泛的酶.
为酶研究和抑制剂查提供了一个有前途的工具.

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Enzyme-Linked Immunosorbent Assay

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or...