Department of Chemical and Biological Engineering and Department of Biomolecular Chemistry, University of Wisconsin, Madison, Wisconsin 53706, USA.
ACS Appl Mater Interfaces. 2010 Mar;2(3):722-31. doi: 10.1021/am900753v.
Surface-induced ordering of liquid crystals (LCs) offers the basis of a label-free analytical technique for the detection of surface-bound biomolecules. The orientation-dependent energy of interaction of a LC with a surface (anchoring energy of LC), in particular, is both sensitive to the presence of surface-bound molecules and easily quantified. Herein, we report a study that analyzes a simple model of twisted nematic LC systems and thereby identifies surfaces with LC anchoring energies in the range of 0.5 microJ/m(2) to 2.0 microJ/m(2) to be optimal for use with LC-based analytical methods. Guided by these predictions, we demonstrate that analytic surfaces possessing anchoring energies within this range can be fabricated with a high level of precision (< 0.1 microJ/m(2)) through formation of monolayers of organothiols (with omega-functional groups corresponding to oligoethyleneglycols and amines) on gold films deposited by physical vapor deposition at oblique angles of incidence. Finally, by using the human epidermal growth factor receptor (EGFR) as a model protein analyte, we have characterized the influence of the anchoring energies of the surfaces on the response of the LC to the presence of surface-bound EGFR. These results, when combined with (32)P-radiolabeling of the EGFR to independently quantify the surface concentration of EGFR, permit identification of surfaces that allow use of LCs to report surface densities of EGFR of 30-40 pg/mm(2). Overall, the results reported in this paper guide the design of surfaces for use in LC-based analytical systems.
表面诱导的液晶(LC)有序提供了一种无标记分析技术的基础,可用于检测表面结合的生物分子。LC 与表面(LC 的锚定能)之间的相互作用的取向依赖性能量,特别是对表面结合分子的存在敏感,并且易于定量。在此,我们报告了一项分析扭曲向列型 LC 系统的简单模型的研究,从而确定了具有 0.5 μJ/m(2)至 2.0 μJ/m(2)范围内的 LC 锚定能的表面对于基于 LC 的分析方法是最佳的。受这些预测的指导,我们证明了通过在物理气相沉积(以倾斜入射角)沉积的金膜上形成有机硫醇(具有对应于聚乙二醇和胺的 ω-官能团)的单层,可以以高精度(<0.1 μJ/m(2)) 制造具有该范围内的锚定能的分析表面。最后,通过使用人表皮生长因子受体(EGFR)作为模型蛋白分析物,我们已经表征了表面的锚定能对 LC 对表面结合的 EGFR 的响应的影响。将这些结果与 EGFR 的(32)P-放射性标记相结合,以独立定量 EGFR 的表面浓度,可识别允许使用 LC 报告 EGFR 的表面密度为 30-40 pg/mm(2)的表面。总的来说,本文报道的结果指导了用于基于 LC 的分析系统的表面设计。