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基于 Zr(4+)-功能化介孔 SiO2 微球的流式细胞术辅助混合读取法用于超灵敏检测蛋白激酶活性

Flow cytometry-assisted mix-and-read assay for ultrasensitive detection of protein kinase activity by use of Zr(4+)-functionalized mesoporous SiO2 microspheres.

机构信息

Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, and Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University , Xi'an 710062, Shaanxi Province, People's Republic of China.

出版信息

Anal Chem. 2013 Nov 19;85(22):10956-61. doi: 10.1021/ac4024457. Epub 2013 Nov 4.

Abstract

A flow cytometry-assisted mix-and-read assay is developed for ultrasensitive detection of protein kinase activity by use of Zr(4+)-functionalized mesoporous SiO2 microspheres (ZrMMs). This strategy integrates the distinct advantages of ZrMMs for highly specific recognition as well as high capacity binding of kinase-induced fluorescent phosphopeptides and flow cytometry for powerful and separation-free bead analysis, leading to an ultrahigh sensitivity for kinase analysis in a extremely simple mix-and-read manner. Furthermore, this ultrasensitive design is well suitable for detection of cell kinase activities in complex biological samples and for screening of potential protein kinase inhibitors, which is of great significance for the development of targeted therapy, clinical diagnosis, and studies of cellular signal transduction pathways.

摘要

一种基于流式细胞术的混合读取检测方法,通过使用 Zr(4+)-功能化介孔二氧化硅微球(ZrMMs),实现对蛋白激酶活性的超灵敏检测。该策略整合了 ZrMMs 作为高度特异性识别以及对激酶诱导的荧光磷酸肽的高容量结合的独特优势,以及流式细胞术对强大且无需分离的珠分析的优势,从而以极其简单的混合读取方式实现了对激酶分析的超高灵敏度。此外,这种超灵敏设计非常适合于复杂生物样品中细胞激酶活性的检测以及潜在蛋白激酶抑制剂的筛选,对于靶向治疗、临床诊断和细胞信号转导途径的研究具有重要意义。

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