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首页> 外文期刊>Annual Review of Analytical Chemistry >Functionalizing Microporous Membranes for Protein Purification and Protein Digestion
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Functionalizing Microporous Membranes for Protein Purification and Protein Digestion

机译：功能化微孔膜用于蛋白质纯化和蛋白质消化

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摘要

This review examines advances in the functionalization of microporous membranes for protein purification and the development of protease-containing membranes for controlled protein digestion prior to mass spectrometry analysis. Recent studies confirm that membranes are superior to bead-based columns for rapid protein capture, presumably because convective mass transport in membrane pores rapidly brings proteins to binding sites. Modification of porous membranes with functional polymeric films or TiO2 nanoparticles yields materials that selectively capture species ranging from phosphopeptides to His-tagged proteins, and protein-binding capacities often exceed those of commercial beads. Thin membranes also provide a convenient framework for creating enzyme-containing reactors that afford control over residence times. With millisecond residence times, reactors with immobilized proteases limit protein digestion to increase sequence coverage in mass spectrometry analysis and facilitate elucidation of protein structures. This review emphasizes the advantages of membrane-based techniques and concludes with some challenges for their practical application.
机译：这篇综述检查了质谱分析之前微孔膜功能化的进展以及用于蛋白质纯化的含蛋白酶膜的发展，以控制蛋白质的消化。最近的研究证实，在快速蛋白质捕获方面，膜优于基于微珠的色谱柱，大概是因为膜孔中的对流传质迅速将蛋白质带到了结合位点。用功能性聚合物膜或TiO2纳米颗粒对多孔膜进行改性，可产生可选择性捕获从磷酸肽到带有His标签的蛋白质的物质，并且蛋白质结合能力通常超过商业珠子。薄膜还为建立含酶反应器提供了方便的框架，该反应器可控制停留时间。具有毫秒级的停留时间，带有固定蛋白酶的反应器会限制蛋白质的消化，从而增加质谱分析中的序列覆盖率，并有助于阐明蛋白质结构。这篇综述强调了基于膜的技术的优势，并以其实际应用面临一些挑战。

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