用于双相环境中筛选反应的facs -可选三乳液微反应器

IF 4.3 3区 材料科学 Q2 CHEMISTRY, MULTIDISCIPLINARY
Samuel Thompson, Yanrong Zhang, Zijian Yang, Lisa Nichols, Polly M. Fordyce
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引用次数: 0

摘要

双相环境可以使任何单一溶剂导致底物溶解度差或催化剂反应性差的化学反应成功。为了高通量筛选双相反应,基于微流控双乳的平台可以使用广泛使用的FACS(荧光活化细胞分选)机器在几个小时内筛选数百万皮升的反应器。然而,将两相反应封装在双乳液中以形成facs可分选的液滴微反应器需要优化溶剂相和表面活性剂,以产生在数小时试验中稳定且与所需反应条件相容的三乳液液滴。这项工作展示了这种具有氟碳外壳和双相辛醇水芯的可分型三乳液微反应器。首先,筛选表面活性剂以稳定用于微反应器堆芯的水包辛醇乳液。在此优化条件下,制备了稳定的三乳微反应器(70%的液滴存活至24小时),通过无细胞蛋白合成在双相核心中产生蛋白质,并使用商用FACS分选机在100 Hz下对三乳进行荧光分选,液滴回收率为75-80%。最后,一个液滴内脂肪酶试验与荧光再间苯二酚底物分割成辛醇被证明。这些三重乳液微反应器在未来有潜力筛选珠状编码的催化剂库,包括用于生物燃料生产的酶,如脂肪酶。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

FACS-Sortable Triple Emulsion Picoreactors for Screening Reactions in Biphasic Environments

FACS-Sortable Triple Emulsion Picoreactors for Screening Reactions in Biphasic Environments

Biphasic environments can enable successful chemical reactions where any single solvent results in poor substrate solubility or poor catalyst reactivity. For screening biphasic reactions at high throughput, a platform based on microfluidic double emulsions can use widely available FACS (Fluorescence Activated Cell Sorting) machines to screen millions of picoliter reactors in a few hours. However, encapsulating biphasic reactions within double emulsions to form FACS-sortable droplet picoreactors requires optimized solvent phases and surfactants to produce triple emulsion droplets that are stable over multi-hour assays and compatible with desired reaction conditions. This work demonstrates such FACS-sortable triple emulsion picoreactors with a fluorocarbon shell and biphasic octanol-in-water core. First, surfactants are screened to stabilize octanol-in-water emulsions for the picoreactor core. With these optimized conditions, stable triple emulsion picoreactors (>70% of droplets survived to 24 hr), produced protein in the biphasic core via cell-free protein synthesis are generated, and sorted these triple emulsions based on fluorescence using a commercial FACS sorter at >100 Hz with 75–80% of droplets recovered. Finally, an in-droplet lipase assay with a fluorogenic resorufin substrate that partitions into octanol is demonstrated. These triple emulsion picoreactors have the potential for future screening bead-encoded catalyst libraries, including enzymes such as lipases for biofuel production.

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来源期刊
Advanced Materials Interfaces
Advanced Materials Interfaces CHEMISTRY, MULTIDISCIPLINARY-MATERIALS SCIENCE, MULTIDISCIPLINARY
CiteScore
8.40
自引率
5.60%
发文量
1174
审稿时长
1.3 months
期刊介绍: Advanced Materials Interfaces publishes top-level research on interface technologies and effects. Considering any interface formed between solids, liquids, and gases, the journal ensures an interdisciplinary blend of physics, chemistry, materials science, and life sciences. Advanced Materials Interfaces was launched in 2014 and received an Impact Factor of 4.834 in 2018. The scope of Advanced Materials Interfaces is dedicated to interfaces and surfaces that play an essential role in virtually all materials and devices. Physics, chemistry, materials science and life sciences blend to encourage new, cross-pollinating ideas, which will drive forward our understanding of the processes at the interface. Advanced Materials Interfaces covers all topics in interface-related research: Oil / water separation, Applications of nanostructured materials, 2D materials and heterostructures, Surfaces and interfaces in organic electronic devices, Catalysis and membranes, Self-assembly and nanopatterned surfaces, Composite and coating materials, Biointerfaces for technical and medical applications. Advanced Materials Interfaces provides a forum for topics on surface and interface science with a wide choice of formats: Reviews, Full Papers, and Communications, as well as Progress Reports and Research News.
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