Microfluidic approaches for the analysis of protein-protein interactionsin solution.
Microfluidic approaches for the analysis of protein-protein interactionsin solution.
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DOI：
10.1007/s12551-020-00679-4
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发表时间：
2020-04-01
影响因子：
--
通讯作者：
Knowles, Tuomas P J
中科院分区：
文献类型：
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作者：
Arter, William E;Levin, Aviad;Knowles, Tuomas P J
人类蛋白质组的探索和表征是一个关键目标，可以加深对生物功能、故障和药物设计的理解。由于蛋白质通常通过与其他蛋白质结合来展示其行为，因此探测蛋白质-蛋白质相互作用的挑战一直是新的和改进的实验方法的焦点。在这里，我们回顾了最近开发的用于研究和量化蛋白质-蛋白质相互作用的微流体技术。我们专注于利用微流体的固有强度来控制微米级质量传输的方法，以促进相互作用蛋白质的无表面和无膜询问和定量。因此，这里描述的微流体工具提供了深入了解生理条件下蛋白质-蛋白质相互作用的能力。我们首先讨论微流体的定义原理，以及利用微尺度流体的扩散控制混合特性来分析蛋白质-蛋白质相互作用的方法。然后，我们描述了利用电泳力来操纵和分级相互作用的蛋白质系统以进行生物物理表征的技术，然后讨论使用微滴区室化来分析蛋白质相互作用的策略。最后，我们强调了该领域的未来方向，例如将微流体实验集成到高通量工作流程中以研究蛋白质相互作用网络。
Exploration and characterisation of the human proteome is a key objective enabling a heightened understanding of biological function, malfunction and pharmaceutical design. Since proteins typicallyexhibit their behaviourby bindingto other proteins, the challenge of probing protein-protein interactions has been the focus of new and improved experimental approaches. Here, we review recently developed microfluidic techniques for the study and quantification of protein-protein interactions. We focus on methodologies that utilise the inherent strength of microfluidics for the control of mass transport on the micron scale, to facilitate surface and membrane-free interrogation and quantification of interacting proteins. Thus, the microfluidic tools described here provide the capability to yield insights on protein-protein interactions under physiological conditions. We first discussthe defining principles of microfluidics, and methods for the analysis of protein-protein interactions that utilise the diffusion-controlled mixing characteristic of fluids at the microscale. We then describetechniques that employ electrophoretic forces to manipulate and fractionate interacting protein systems for their biophysical characterisation, before discussing strategies that use microdroplet compartmentalisation for the analysis of protein interactions. We conclude by highlighting future directions for the field, such as the integration of microfluidic experiments into high-throughput workflows for the investigation of protein interaction networks.