微流控装置开发的PDMS生物功能化研究:唾液皮质醇的应用

V. Pinto, G. Minas, M. Correia-Neves
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引用次数: 3

摘要

本文介绍了PDMS(聚二甲基硅氧烷)表面功能化的研究,以开发一种微流控免疫传感器,通过光学检测定量分析唾液皮质醇。采用不同的抗体固定方法在PDMS表面实现功能化:(a)被动吸附固定在原始PDMS上;(b)用(3-氨基丙基)-三乙氧基硅烷(APTES)对PDMS表面进行硅烷化生成氨基,并用交联剂戊二醛(GA)将抗体后共价固定在APTES-PDMS上;(c)在PDMS表面涂覆BSA,阻断非特异性蛋白吸附,再通过GA使蛋白A共价键。在最后一种方法中,抗体被共价固定在蛋白A上,因为它与抗体的恒定分数区(Fc)有很高的亲和力。原子力显微镜(AFM)和分光光度分析表明,蛋白A的固定化方法具有更高的效率,可以获得更高的表面粗糙度和均匀性,并获得更高的吸光度信号。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
PDMS biofunctionalization study for the development of a microfluidic device: Application to salivary cortisol
This paper presents the study of the PDMS (poly(dimethylsiloxane)) surface functionalization for the development of a microfluidic immunosensor that quantitatively analyss salivary cortisol by optical detection. The functionalization was performed using different antibodies immobilization methods on PDMS surface: (a) immobilization by passive adsorption on pristine PDMS; (b) silanization of PDMS surface with (3-aminopropyl)-triethoxysilane (APTES) to generate amino groups and posterior covalent immobilization of antibodies on APTES-PDMS using cross-linker glutaraldehyde (GA); (c) coating the PDMS surface with BSA to block nonspecific protein adsorption, and then covalent bond of the protein A via GA. In this last approach, the antibodies were covalently immobilized to protein A due to its high affinity with the constant fraction (Fc) region of the antibodies. Atomic force microscope (AFM) and spectrophotometric analysis demonstrated that the immobilization method using protein A is more efficient since a higher roughness and uniformity on the PDMS surface and higher absorbance signals were obtained.
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