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染色蛋白质:提高吸附研究中椭圆测量灵敏度的简单方法。

Staining proteins: a simple method to increase the sensitivity of ellipsometric measurements in adsorption studies.

机构信息

Department of Chemistry, The University of Texas at San Antonio, One UTSA Circle, San Antonio, TX 78249, USA.

出版信息

Colloids Surf B Biointerfaces. 2011 Jan 1;82(1):253-7. doi: 10.1016/j.colsurfb.2010.08.021. Epub 2010 Aug 21.

Abstract

This communication describes a simple way to improve the sensitivity of spectroscopic ellipsometry, when applied to monitor the adsorption of proteins to solid surfaces. The method described herein is based on the reaction of a commercially available dye (Coomassie brilliant blue G) with the adsorbed proteins and the subsequent analysis by spectroscopic ellipsometry. In order to demonstrate the potential advantages of this method, the adsorption of bovine serum albumin to an antifouling coating was also investigated. According to our results, the modification with the dye significantly affects the optical properties of the adsorbed protein layer, which can be represented using a simple optical model (Lorentz). In general, the proposed modification increases the sensitivity of the detection by 2.5 ± 0.4-fold and enables the analysis of thin layers of adsorbed protein not obtainable by conventional methods. These results particularly reveal the importance of the proposed modification for the evaluation of low adsorbing substrates and antifouling coatings.

摘要

本通讯介绍了一种提高光谱椭圆偏振术灵敏度的简单方法,该方法可用于监测蛋白质在固体表面上的吸附。本文所描述的方法基于一种市售染料(考马斯亮蓝 G)与吸附蛋白质的反应,以及随后通过光谱椭圆偏振术进行的分析。为了证明该方法的潜在优势,还研究了牛血清白蛋白在防污涂层上的吸附。根据我们的结果,染料的修饰显著影响了吸附蛋白质层的光学性质,这可以用简单的光学模型(洛伦兹)来表示。一般来说,所提出的修饰将检测灵敏度提高了 2.5±0.4 倍,并使通过常规方法无法获得的薄吸附蛋白质层的分析成为可能。这些结果特别揭示了所提出的修饰对于评估低吸附底物和防污涂层的重要性。

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