Sorting by interfacial tension (SIFT): Label-free enzyme sorting using droplet microfluidics
Sorting by interfacial tension (SIFT): Label-free enzyme sorting using droplet microfluidics
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DOI：
10.1016/j.aca.2019.08.025
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发表时间：
2019-12-16
影响因子：
6.2
通讯作者：
Abbyad, Paul
中科院分区：
文献类型：
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作者：
Horvath, Daniel G.;Braza, Samuel;Abbyad, Paul
液滴微流体具有通过在惰性油中流动的皮升液滴中携带试剂来大大增加酶的筛选和分选的通量的能力。人们发现，通过使用特定的表面活性剂，液滴的界面张力对液滴pH值非常敏感。当受限的液滴遇到微加工的沟槽时，这使得能够对不同pH值的液滴进行分选。该设备可以扩展到排序酶，因为大量的酶促反应导致酸性或碱性产物的产生以及溶液pH值的同时变化。酶促反应的进展从液滴的流动序列的位置被跟踪。我们展示了酯酶同工酶的分类根据其酶活性。这种无标记的技术，我们称之为液滴分选界面张力（SIFT），不需要活性成分，并将在高通量应用，包括酶筛选和酶的定向进化酶分选的应用。(C)2019爱思唯尔B.V.保留所有权利。
Droplet microfluidics has the ability to greatly increase the throughput of screening and sorting of enzymes by carrying reagents in picoliter droplets flowing in inert oils. It was found with the use of a specific surfactant, the interfacial tension of droplets can be very sensitive to droplet pH. This enables the sorting of droplets of different pH when confined droplets encounter a microfabricated trench. The device can be extended to sort enzymes, as a large number of enzymatic reactions lead to the production of an acidic or basic product and a concurrent change in solution pH. The progress of an enzymatic reaction is tracked from the position of a flowing train of droplets. We demonstrate the sorting of esterase isoenzymes based on their enzymatic activity. This label-free technology, that we dub droplet sorting by interfacial tension (SIFT), requires no active components and would have applications for enzyme sorting in high-throughput applications that include enzyme screening and directed evolution of enzymes. (C) 2019 Elsevier B.V. All rights reserved.