Xue Chang-Ying, Yang Kun-Lin
Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117576.
Langmuir. 2007 May 8;23(10):5831-5. doi: 10.1021/la070076+. Epub 2007 Apr 11.
In this paper we present a study of using oxygen plasma for chemically modifying inert hydrocarbon self-assembled monolayers of octadecyltrichlorosilane (OTS-SAMs) and rendering active surfaces for protein immobilization. Detailed surface modification and protein immobilization were characterized by using ellipsometry, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared-attenuated total reflectance spectroscopy, and fluorescence microscopy. Our XPS results showed that the surface reaction between OTS-SAMs and oxygen plasma can generate new surface functional groups such as alcohol (C-O), aldehyde (C=O), and carboxylic acid (O-C=O), and their compositions can be controlled by using different treatment times and powers. A short treatment time ( approximately 1 s) and high power (10 W) can lead to a higher density of aldehyde groups, which can serve as linker groups for protein immobilization through the formation of Schiff bases with the amine groups of proteins. By using the fluorescence immunostaining method, we confirmed that human immunoglobulin (IgG) can be immobilized on a glass slide, only if the surface was decorated with OTS-SAMs and if the OTS-SAMs were pretreated with oxygen plasma. The protein immobilized on the oxygen-plasma-treated surface can only be recognized by using a highly specific antibody, FITC-anti-IgG, but not FITC-anti-biotin.
在本文中,我们展示了一项关于使用氧等离子体对十八烷基三氯硅烷(OTS - 自组装单分子层)的惰性碳氢化合物自组装单分子层进行化学改性并使表面具有活性以用于蛋白质固定的研究。通过椭偏仪、X射线光电子能谱(XPS)、傅里叶变换红外衰减全反射光谱和荧光显微镜对详细的表面改性和蛋白质固定进行了表征。我们的XPS结果表明,OTS - 自组装单分子层与氧等离子体之间的表面反应可以产生新的表面官能团,如醇(C - O)、醛(C = O)和羧酸(O - C = O),并且它们的组成可以通过使用不同的处理时间和功率来控制。较短的处理时间(约1秒)和高功率(10瓦)可导致醛基密度更高,醛基可作为连接基团,通过与蛋白质的胺基形成席夫碱来实现蛋白质固定。通过荧光免疫染色方法,我们证实只有当表面用OTS - 自组装单分子层修饰且OTS - 自组装单分子层经过氧等离子体预处理时,人免疫球蛋白(IgG)才能固定在载玻片上。固定在氧等离子体处理过的表面上的蛋白质只能被高特异性抗体FITC - 抗IgG识别,而不能被FITC - 抗生物素识别。