Department of Mechanical Engineering, KAIST , 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Korea.
Department of Proteome Structural Biology, KRIBB School of Bioscience, Korea University of Science and Technology , 125 Gwahak-ro Yuseong-gu, Daejeon 34141, Korea.
Anal Chem. 2017 Dec 19;89(24):13313-13319. doi: 10.1021/acs.analchem.7b03474. Epub 2017 Nov 30.
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.
我们开发了一种混合微流控装置,利用声波在连续流微通道内驱动功能化的微颗粒,并从复杂流体中分离出与颗粒结合的靶蛋白。该声流控装置由叉指换能器组成,叉指换能器产生高频表面声波(SAW)和聚二甲基硅氧烷(PDMS)微流道。SAW 与微通道内的样品流体相互作用,使颗粒偏离其原始流线,从而实现分离。链霉亲和素功能化的聚苯乙烯(PS)微球用于捕获一端标记有生物素分子的适体(单链 DNA)。通过链霉亲和素-生物素连接固定在微球上的定制适体 15(apt15)的自由端,与凝血酶(th)的外位结合,形成 PS-apt15-th 复合物,用于捕获模型靶蛋白凝血酶。我们证明 PS-apt15 缀合物在复杂流体中选择性地捕获凝血酶分子。形成 PS-apt15-th 复合物后,将样品流体与两种缓冲鞘流一起泵入微通道,在 SAW 暴露之前,缓冲鞘流通过流体动力学聚焦样品流,以实现 PS-apt15-th 从非靶蛋白中的分离。我们使用所提出的声流控装置成功地从 mCardinal2 和人血清中分离出血栓酶。