Microfluidic Diffusion Analysis of the Sizes and Interactions of Proteins under Native Solution Conditions
Microfluidic Diffusion Analysis of the Sizes and Interactions of Proteins under Native Solution Conditions
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DOI：
10.1021/acsnano.5b04713
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发表时间：
2016-01-01
期刊：
影响因子：
17.1
通讯作者：
Knowles, Tuomas P. J.
中科院分区：
文献类型：
--
作者：
Arosio, Paolo;Mueller, Thomas;Knowles, Tuomas P. J.
在自然条件下表征大分子的大小和相互作用是分子科学的许多领域中的一个具有挑战性的问题，这从根本上源于生物分子混合物的多分散性。在这里，我们描述了一个微流控平台的基础上监测微米级的质量传输同时在空间和时间的扩散尺寸。我们表明，这种组合的时空数据的全球分析，使混合物中的单个物种的流体动力学半径直接通过去卷积平均信号到单个物种的贡献。我们证明，进行快速非侵入性大小的能力，允许这种方法被用来表征在自然条件下的生物分子之间的相互作用。我们通过实施单步定量免疫测定来说明该技术的潜力，该免疫测定在秒的时间尺度上操作，并检测复杂混合物中生物分子之间的特定相互作用。
Characterizing the sizes and interactions of macromolecules under native conditions is a challenging problem in many areas of molecular sciences, which fundamentally arises from the polydisperse nature of biomolecular mixtures. Here, we describe a microfluidic platform for diffusional sizing based on monitoring micron-scale mass transport simultaneously in space and time. We show that the global analysis of such combined space time data enables the hydrodynamic radii of individual species within mixtures to be determined directly by deconvoluting average signals into the contributions from the individual species. We demonstrate that the ability to perform rapid noninvasive sizing allows this method to be used to characterize interactions between biomolecules under native conditions. We illustrate the potential of the technique by implementing a single-step quantitative immunoassay that operates on a time scale of seconds and detects specific interactions between biomolecules within complex mixtures.