Department of Chemistry, University of Illinois at Urbana-Champaign , 600 South Mathews Avenue, Urbana, Illinois 61801, United States.
ACS Appl Mater Interfaces. 2011 Sep;3(9):3762-71. doi: 10.1021/am2009597. Epub 2011 Aug 12.
Methods for the generation of substratespresenting biomolecules in a spatially controlled manner are enabling tools for applications in biosensor systems, microarray technologies, fundamental biological studies and biointerface science. We have implemented a method to create biomolecular patterns by using light to control the direct covalent immobilization of biomolecules onto benzophenone-modified glass substrates. We have generated substrates presenting up to three different biomolecules patterned in sequence, and demonstrate biomolecular photopatterning on corrugated substrates. The chemistry of the underlying monolayer was optimized to incorporate poly(ethylene glycol) to enable adhesive cell adhesion onto patterned extracellular matrix proteins. Substrates were characterized with contact angle goniometry, AFM, and immunofluorescence microscopy. Importantly, radioimmunoassays were performed to quantify the site density of immobilized biomolecules on photopatterned substrates. Retained function of photopatterned proteins was demonstrated both by native ligand recognition and cell adhesion to photopatterned substrates, revealing that substrates generated with this method are suitable for probing specific cell receptor-ligand interactions. This molecularly general photochemical patterning method is an enabling tool for the creation of substrates presenting both biochemical and topographical variation, which is an important feature of many native biointerfaces.
用于以空间控制方式呈现生物分子的底物的生成方法是在生物传感器系统、微阵列技术、基础生物学研究和生物界面科学中应用的有效工具。我们已经实施了一种通过使用光来控制生物分子直接共价固定到苯并二酮修饰的玻璃基底上的方法,从而生成生物分子图案。我们已经生成了呈现多达三种不同生物分子的顺序图案的底物,并在波纹状基底上演示了生物分子光图案化。优化了底层单层的化学性质以纳入聚(乙二醇),从而能够将细胞粘附到图案化的细胞外基质蛋白上。使用接触角测角法、AFM 和免疫荧光显微镜对底物进行了表征。重要的是,进行放射免疫测定以定量光图案化底物上固定化生物分子的位点密度。通过天然配体识别和细胞粘附到光图案化的基底上来证明光图案化蛋白的保留功能,这表明使用该方法生成的基底适用于探测特定的细胞受体-配体相互作用。这种分子通用的光化学图案化方法是创建呈现生化和形貌变化的底物的有效工具,这是许多天然生物界面的重要特征。