In-situ protein adsorption study on biofunctionalized surfaces using spectroscopic ellipsometry
In-situ protein adsorption study on biofunctionalized surfaces using spectroscopic ellipsometry
复制标题
DOI：
10.1016/j.tsf.2009.10.110
复制
发表时间：
2010-02-01
期刊：
影响因子：
2.1
通讯作者：
Subramanian, Anuradha
中科院分区：
文献类型：
--
作者：
Goyal, Dileep K.;Subramanian, Anuradha
目前用于评估表面上生物分子相互作用的技术需要使用放射性标记、酶或荧光标记来记录和报告结合事件。椭圆偏振法已被证明是理解固体基质上生物分子相互作用的有力工具，并且通常不需要标记配体或受体。在本研究中，人血清白蛋白（HSA）的功能化硅表面上的吸附动力学进行了评估，使用原位椭圆偏振。原位椭圆偏振法被用来估计吸附层的厚度和HSA的吸附和解吸动力学功能化表面上。在这项研究中，使用氨丙基三乙氧基硅烷（APTES）以及使用APTES和甲基三乙氧基硅烷以1：10的比例混合的硅烷制备了致密的自组装单层，作为蛋白质固定在硅表面的模板。使用三种不同的配体/受体进一步修饰硅烷衍生表面，据报道这些配体/受体可结合HSA，即：线性肽、抗人血清白蛋白的多克隆抗体和小合成配体（2，4，6-三（二甲基氨基甲基）苯酚）。观察到吸附的HSA的量随时间和HSA溶液的初始浓度而增加。HSA在功能化表面上的吸附动力学近似于蛋白质吸附的简单模型。一个很好的模型拟合的实验数据，从而使功能化的硅表面上的HSA的吸附动力学的解释。研究了不同的HSA结合配体对蛋白质吸附和解吸速率常数的影响。由爱思唯尔公司出版
Techniques currently employed to evaluate biomolecular interactions on surfaces require the use of radiolabeled, enzymatic, or fluorescent-tags to record and report the binding event. Ellipsometry has proven to be a powerful tool in understanding the biomolecular interactions on solid substrates and, typically does not require the labeling of the ligand or the receptor. In this present study, the adsorption kinetics of Human Serum Albumin (HSA) on functionalized silicon surfaces were evaluated using in-situ ellipsometry. In-situ ellipsometry was used to estimate the thickness of the adsorbed layers and the adsorption and desorption kinetics of HSA on functionalized surfaces. In this study, dense, self assembled monolayers were fabricated using aminopropyltriethoxysilane (APTES) and mixed silanes using APTES and methyltriethoxysilane at a ratio of 1:10, to serve as a template for protein immobilization on silicon surfaces. The silane derivatized surfaces were further modified using three different ligands/receptors that have been reported to bind HSA, namely: a linear peptide, a polyclonal antibody against human serum albumin, and small synthetic ligand (2, 4, 6-Tris(dimethylaminomethyl) phenol. The amount of HSA adsorbed was observed to increase with time, and with the initial concentration of the HSA solution. The adsorption kinetics of HSA on functionalized surfaces was approximated by a simple model for Protein adsorption. A good model fit was obtained for the experimental data, thus enabling the interpretation of the adsorption kinetics of HSA on functionalized silicon surfaces. The effect of different HSA binding ligands on the rate constants affecting protein adsorption and desorption were studied. Published by Elsevier B.V.