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Acoustic Wave-Driven Functionalized Particles for Aptamer-Based Target Biomolecule Separation

机译:基于Aptamer的靶向生物分子分离的声波驱动官能化颗粒

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摘要

We developed a hybrid microfluidic device that utilized acoustic waves to drive functionalized microparticles inside a continuous flow microchannel and to separate particle-conjugated target proteins from a complex fluid. The acoustofluidic device is composed of an interdigitated transducer that produces high-frequency surface acoustic waves (SAW) and a polydimethylsiloxane (PDMS) microfluidic channel. The SAW interacted with the sample fluid inside the microchannel and deflected particles from their original streamlines to achieve separation. Streptavidin-functionalized polystyrene (PS) microparticles were used to capture aptamer (single-stranded DNA) labeled at one end with a biotin molecule. The free end of the customized aptamer15 (apt15), which was attached to the microparticles via streptavidin-biotin linkage to form the PS-apt15 conjugate, was used to capture the model target protein, thrombin (th), by binding at exosite I to form the PS-apt15-th complex. We demonstrated that the PS-apt15 conjugate selectively captured thrombin molecules in a complex fluid. After the PS-apt15-th complex was formed, the sample fluid was pumped through a PDMS microchannel along with two buffer sheath flows that hydrodynamically focused the sample flow prior to SAW exposure for PS-apt15-th separation from the non-target proteins. We successfully separated thrombin from mCardinal2 and human serum using the proposed acoustofluidic device.
机译:我们开发了一种混合微流体装置,其利用声波在连续流动微通道内驱动功能化微粒,并将颗粒缀合的靶蛋白与复杂的流体分离。声副流体装置由互频换能器组成,其产生高频表面声波(SAW)和聚二甲基硅氧烷(PDMS)微流体通道。锯与微通道内的样品流体与其原始流线中的偏转颗粒相互作用以实现分离。用生物素分子使用链霉抗生物素蛋白官能化聚苯乙烯(PS)微粒捕获标记的适体(单链DNA)。通过链霉抗生物素蛋白 - 生物素键连接的定制Aptamer15(APT15)的自由端用于形成PS-APT15缀合物,用于通过在后化I中结合捕获模型靶蛋白凝血酶(TH)形成PS-APT15-TH复合物。我们证明了PS-APT15缀合物在复杂的流体中选择性地捕获凝血酶分子。在形成PS-APT15-TH复合物之后,将样品流体通过PDMS微通道泵送,与两个缓冲鞘流一起,其在从非靶蛋白看到PS-APT15分离之前,进一步地将样动力学聚焦的样品流动。我们使用所提出的声毒流体装置成功地从McArdinal2和人血清分离出凝血酶。

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  • 来源
    《Analytical chemistry》 |2017年第24期|共7页
  • 作者单位

    Korea Adv Inst Sci &

    Technol Dept Mech Engn 291 Daehak Ro Daejeon 34141 South Korea;

    Korea Adv Inst Sci &

    Technol Dept Mech Engn 291 Daehak Ro Daejeon 34141 South Korea;

    Korea Univ Sci &

    Technol KRIBB Sch Biosci Dept Proteome Struct Biol 125 Gwahak Ro Daejeon 34141 South Korea;

    Korea Adv Inst Sci &

    Technol Dept Mech Engn 291 Daehak Ro Daejeon 34141 South Korea;

    Korea Adv Inst Sci &

    Technol Dept Mech Engn 291 Daehak Ro Daejeon 34141 South Korea;

    Korea Adv Inst Sci &

    Technol Dept Mech Engn 291 Daehak Ro Daejeon 34141 South Korea;

    Korea Adv Inst Sci &

    Technol Dept Mech Engn 291 Daehak Ro Daejeon 34141 South Korea;

    Korea Univ Sci &

    Technol KRIBB Sch Biosci Dept Proteome Struct Biol 125 Gwahak Ro Daejeon 34141 South Korea;

    Korea Adv Inst Sci &

    Technol Dept Mech Engn 291 Daehak Ro Daejeon 34141 South Korea;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
  • 关键词

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