Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, Wisconsin 53706, USA.
Langmuir. 2009 Oct 20;25(20):12275-82. doi: 10.1021/la9017135.
The ability to pattern small molecules and proteins on artificial surfaces is of importance for the development of new tools including tissue engineering, cell-based drug screening, and cell-based sensors. We describe here a novel "caged" thiol-mediated strategy for the fabrication of planar substrates patterned with biomolecules using photolithography. A thiol-bearing phosphoramidite (3-(2'-nitrobenzyl)thiopropyl (NBTP) phosphoramidite) was synthesized and coupled to a hydroxyl-terminated amorphous carbon substrate. A biocompatible oligo(ethylene glycol) spacer was used to resist nonspecific adsorption of protein and DNA and enhance flexibility of attached biomolecules. Thiol functionalities are revealed by UV irradiation of NBTP-modified surfaces. Both the surface coupling and photodeprotection were monitored by Polarization Modulation Fourier Transform Infrared Reflection Absorption Spectroscopy (PM-FTIRRAS) and X-ray Photoelectron Spectroscopy (XPS) measurements. The newly exposed thiols are chemically very active and react readily with a wide variety of groups. A series of molecules including biotin, DNA, and proteins were attached to the surfaces with retention of their biological activities, demonstrating the utility and generality of the approach.
在人工表面上对小分子和蛋白质进行图案化的能力对于开发新工具非常重要，包括组织工程、基于细胞的药物筛选和基于细胞的传感器。我们在这里描述了一种新的“笼状”硫醇介导的策略，用于使用光刻法制造具有生物分子的平面基底的图案化。合成了带有巯基的磷酰胺（3-(2'-硝基苄基)巯基丙基（NBTP）磷酰胺）并偶联到羟基封端的非晶碳基底上。使用生物相容性的聚（乙二醇）间隔物来抵抗蛋白质和 DNA 的非特异性吸附，并增强附着的生物分子的灵活性。通过 NBTP 修饰表面的紫外线照射来揭示硫醇官能团。通过偏振调制傅里叶变换红外反射吸收光谱（PM-FTIRRAS）和 X 射线光电子能谱（XPS）测量来监测表面偶联和光保护。新暴露的巯基具有非常活泼的化学性质，很容易与各种基团反应。一系列分子，包括生物素、DNA 和蛋白质，被附着到表面上，保留其生物活性，证明了该方法的实用性和通用性。
