微量热泳动技术在生物分子相互作用研究中的应用

doi:10.5246/jcps.2020.09.061

摘要/Abstract

摘要：

生物分子间相互作用的准确表征不仅能够为了解细胞中的生命过程提供必要的知识, 而且还能够为药物的发现和发展铺平道路。生物物理技术由于其在通量和灵敏度上的优势, 已成为研究生物分子相互作用的必不可少的技术之一。而具备低成本、检测速度快、操作体系简单并同时能提供丰富实验数据信息等优势的生物物理技术更是已成为检测生物分子间相互作用的首选技术。在本论文中, 我们总结了使用微量热泳动仪(MicroScale Thermophoresis, MST)进行生物分子间相互作用研究的定性和定量分析成果, 并展示了如何使用MST的拓展功能获得酶动力学参数和蛋白质稳定性参数。作为一种操作简单、结果准确的生物物理技术, MST依据微观温度梯度中分子的热泳移动变化来定量检测亲和力。与其它生物物理技术相比, MST的优势为无需固定、分析时间短、样品消耗少且能够在细胞裂解液等复杂溶液体系中进行亲和力分析等。在本论文中, 我们将依次讨论MST的检测原理、实验操作流程和应用示例。

关键词: 生物分子间相互作用, 生物物理技术, 微量热泳动仪

Abstract:

Accurate characterization of the interactions between biomolecules not only provides fundamental insights into cellular processes but also paves the way for drug discovery and development. With recent increases in throughput and sensitivity, biophysical technologies have become prominent tools for studying biomolecular interactions. Biophysical techniques that can reduce costs, shorten detection time, simplify the complexity of the system under analysis, and simultaneously provide high-quality data content are particularly favored. Here, we summarize the qualitative and quantitative analysis of biomolecular interactions using MicroScale Thermophoresis (MST), as well as extend the application of MST functions to explore thermodynamics, enzyme kinetics and protein folding-unfolding processes. MST has emerged as a simple and powerful biophysical approach for identifying and quantifying binding events based on the movement of molecules along microscopic temperature gradients. The advantages of MST over other competitive biophysical techniques include freedom from immobilization, rapid analysis times, lower sample consumption, and the ability to analyze binding affinities in cell lysates. This article discusses the instrumental setups, principles, experimental workflows, and examples of MST application in practice.

Key words: Biomolecular interactions, Biophysical technologies, MicroScale Thermophoresis

中图分类号:

R917

Supporting:

王倩, 王静, 宋书香, 朱贵旺, 曹泽, 刘振明. 微量热泳动技术在生物分子相互作用研究中的应用[J]. 中国药学(英文版), 2020, 29(9): 656-665.

Qian Wang, Jing Wang, Shuxiang Song, Guiwang Zhu, Ze Cao, Zhenming Liu. Microscale thermophoresis in the investigation of biomolecular interactions[J]. Journal of Chinese Pharmaceutical Sciences, 2020, 29(9): 656-665.

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