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基于二氧化硅表面构建的支撑脂质双层的双偏振干涉和胶束电动色谱分析:其抗蛋白吸附效果的评价。

Dual polarization interferometric and capillary electrophoretic analysis of supported lipid bilayer constructed on silica-based surface: evaluation of its anti-protein adsorption effect.

机构信息

BioAnalytical and Nanobiomedicinal Laboratory, Department of Biochemical Science and Technology, National Taiwan University, Taipei, Taiwan.

出版信息

Anal Chim Acta. 2012 Feb 10;714:127-33. doi: 10.1016/j.aca.2011.11.062. Epub 2011 Dec 7.

Abstract

Supported lipid bilayer (SLB) has been demonstrated as a model of cell membranes with prospective bioanalytical or biotechnological applications. In this study, the formation of SLB and their potential biofunctionality against protein adsorption were investigated by Dual Polarization Interferometry (DPI) and Capillary Electrophoresis (CE). DPI studies on different formulations of double-chained, zwitterionic phospholipidlipids, allow the process of bilayer formation to be followed in situ and in real time. Furthermore the anti-protein adsorption effect provided by the various formulated SLBs was examined by DPI. In addition, the SLB coatings of the same lipid formulations were subsequently employed in CE experiments as a pseudo-stationary phase for demonstrating more efficient separation of alkaline protein standard mixtures. SLB-assisted CE was found to be capable of separating 4 alkaline proteins (protonated at neutral pH). This study demonstrates the applicability of DPI to monitor the process of SLB formation; and our findings, obtained by both DPI and CE, confirm that the presence of the SLB reduced drastically the problematic interactions between cationic, alkaline proteins and the negatively charged silica capillary wall, leading to better recovery and efficient separation of the proteins under investigation.

摘要

支持的脂质双层 (SLB) 已被证明是细胞膜的模型,具有潜在的生物分析或生物技术应用。在这项研究中,通过双折射干涉 (DPI) 和毛细管电泳 (CE) 研究了 SLB 的形成及其对蛋白质吸附的潜在生物功能。DPI 对不同配方的双链、两性离子磷脂的研究允许在原位和实时跟踪双层形成过程。此外,通过 DPI 研究了各种配方 SLB 提供的抗蛋白质吸附效果。此外,相同脂质配方的 SLB 涂层随后在 CE 实验中用作伪固定相,以证明碱性蛋白质标准混合物的分离更有效。SLB 辅助的 CE 被发现能够分离 4 种碱性蛋白质(在中性 pH 下质子化)。这项研究证明了 DPI 可用于监测 SLB 形成过程;并且我们通过 DPI 和 CE 获得的发现证实,SLB 的存在大大减少了阳离子、碱性蛋白质与带负电荷的硅胶毛细管壁之间的问题相互作用,从而导致研究中的蛋白质更好的回收率和有效分离。

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