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Microfluidic chips with multi-junctions: an advanced tool in recovering proteins from inclusion bodies

机译:具有多结点的微流控芯片:从包涵体中回收蛋白质的先进工具

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摘要

Active recombinant proteins are used for studying the biological functions of genes and for the development of therapeutic drugs. Overexpression of recombinant proteins in bacteria often results in the formation of inclusion bodies, which are protein aggregates with non-native conformations. Protein refolding is an important process for obtaining active recombinant proteins from inclusion bodies. However, the conventional refolding method of dialysis or dilution is time-consuming and recovered active protein yields are often low, and a cumbersome trial-and-error process is required to achieve success. To circumvent these difficulties, we used controllable diffusion through laminar flow in microchannels to regulate the denaturant concentration. This method largely aims at reducing protein aggregation during the refolding procedure. This Commentary introduces the principles of the protein refolding method using microfluidic chips and the advantage of our results as a tool for rapid and efficient recovery of active recombinant proteins from inclusion bodies.
机译:活性重组蛋白用于研究基因的生物学功能和开发治疗药物。重组蛋白在细菌中的过表达通常导致包涵体的形成,包涵体是具有非天然构象的蛋白聚集体。蛋白质重折叠是从包涵体获得活性重组蛋白质的重要​​过程。然而,常规的透析或稀释的重折叠方法是费时的,并且回收的活性蛋白产量通常较低,并且需要繁琐的反复试验过程才能获得成功。为了克服这些困难,我们使用了通过微通道中层流的可控扩散来调节变性剂浓度。该方法主要旨在减少重折叠过程中的蛋白质聚集。本评论介绍了使用微流控芯片进行蛋白质复性方法的原理,以及我们的研究结果作为从包涵体中快速有效回收活性重组蛋白的工具的优势。

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