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Lipid-Coated Microdroplet Array for in Vitro Protein Synthesis

机译:脂质包裹的微滴阵列用于体外蛋白质合成

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摘要

ABSTRACT: Monitoring complex biological assays such as in vitro proteinnsynthesis over long periods in micrometer-sized cavities of poly(dimethylnsiloxane) (PDMS) microfluidic devices requires a strategy that solves thenadsorption and absorption problems on PDMS surfaces. In this study, wendeveloped a technique that instantaneously arrays aqueous microdropletsncoated with a phospholipid membrane within a single microfluidic device.nThe simple lipid bilayer coating effectively inhibits the adsorption of proteinsnand DNA, whereas the encapsulation of the droplet reduces the area inncontact with the PDMS surface, resulting in decreased absorption in part.nAlthough the size becomes smaller during the first few hours, a lipid-coated microdroplet array demonstrated a temporal stability ofnmore than 20 h and a size uniformity of CV 3%in the device. Furthermore, we succeeded in expressing a green fluorescent protein bynconfining an in vitro translation system in the microdroplets, which was confirmed by scanning the fluorescence spectrum of thendroplets, demonstrating that the lipid coat secured the synthetic reaction from the adsorption problem.
机译:摘要:监测复杂的生物学检测方法,例如在微米级的聚二甲基硅氧烷(PDMS)流体装置的空腔中进行长时间的体外蛋白质合成,需要一种能够解决PDMS表面吸附和吸收问题的策略。在这项研究中,Wen开发了一种技术,可在单个微流体装置中即时排列涂有磷脂膜的水性微滴。n简单的脂质双层涂层有效抑制蛋白质和DNA的吸附,而微滴的包裹减少了与PDMS表面非接触的面积,尽管最初几小时尺寸变小,但脂质包裹的微滴阵列显示了超过20小时的时间稳定性和CV 3%的尺寸均匀性。此外,我们通过在微滴中建立体外翻译系统成功表达了绿色荧光蛋白,这是通过扫描微滴的荧光光谱证实的,这表明脂质涂层可确保合成反应免受吸附问题的影响。

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  • 来源
    《Analytical Chemistry》 |2011年第8期|p.3186-3191|共6页
  • 作者单位

    †Kanagawa Academy of Science and Technology, Japan‡Laboratory for Integrated Micro and Mechatronic Systems, CNRS-IIS,UMI 2820, The University of Tokyo, Japan§Institute of Industrial Science, The University of Tokyo, Japan;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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