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An off-the-shelf integrated microfluidic device comprising self-assembled monolayers for protein array experiments

机译:包含自组装单分子层的现成集成微流控设备用于蛋白质阵列实验

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摘要

Microfluidic-based protein arrays are promising tools for life sciences, with increased sensitivity and specificity. One of the drawbacks of this technology is the need to create fresh surface chemistry for protein immobilization at the beginning of each experiment. In this work, we attempted to include the process of surface functionalization as part of the fabrication of the device, which would substitute the time consuming step of surface functionalization at the beginning of each protein array experiment. To this end, we employed a novel surface modification using self-assembled monolayers (SAMs) to immobilize biomolecules within the channels of a polydimethylsiloxane (PDMS) integrated microfluidic device. As a model, we present a general method for depositing siloxane-anchored SAMs, with 1-undecyl-thioacetate-trichlorosilane (C11TA) on the silica surfaces. The process involved developing PDMS-compatible conditions for both SAM deposition and functional group activation. We successfully demonstrated the ability to produce, within an integrated microfluidic channel, a C11TA monolayer with a covalently conjugated antibody. The antibody could then bind its antigen with a high signal to background ratio. We further demonstrated that the antibody was still active after storage of the device for a week. Integration of the surface chemistry into the device as part of its fabrication process has potential to significantly simplify and shorten many experimental procedures involving microfluidic–based protein arrays. In turn, this will allow for broader dissemination of this important technology.
机译:基于微流体的蛋白质阵列是具有增强的敏感性和特异性的生命科学的有前途的工具。该技术的缺点之一是需要在每个实验开始时创建新的表面化学来固定蛋白质。在这项工作中,我们尝试将表面功能化过程作为设备制造的一部分,以取代每个蛋白质阵列实验开始时耗时的表面功能化步骤。为此,我们采用了一种使用自组装单分子膜(SAMs)的新型表面改性方法,将生物分子固定在集成的二甲基硅氧烷(PDMS)微流体装置通道内。作为模型,我们介绍了一种在二氧化硅表面上沉积硅氧烷锚定的SAM的一般方法,其中含有1-十一烷基-硫代乙酸酯-三氯硅烷(C11TA)。该过程包括为SAM沉积和官能团激活开发与PDMS兼容的条件。我们成功地证明了在整合的微流体通道内产生具有共价结合抗体的C11TA单层膜的能力。然后,抗体可以高信噪比与抗原结合。我们进一步证明,在将设备存储一周后,抗体仍然具有活性。在设备的制造过程中,将表面化学方法集成到设备中可能会显着简化和缩短许多涉及基于微流体的蛋白质阵列的实验程序。反过来,这将允许更广泛地传播这一重要技术。

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