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首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Bacteria assisted protein imprinting in sol-gel derived films
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Bacteria assisted protein imprinting in sol-gel derived films

机译：细菌辅助蛋白在溶胶 - 凝胶衍生膜中印迹

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A hierarchical imprinting strategy was used to create protein imprints in a silicate film with a high binding capacity as well as selectivity toward the imprint protein and little specificity towards other proteins. In the first part of this work, rod-shaped bacteria were used as templates to create imprints in silica films of various thicknesses to open up the silica framework and increase the surface area exposed to solution. In the second part, the protein (e.g., cytochrome c (CYC) or green fluorescent protein (GFP)) was covalently attached to the surface of Bacillus subtilis and this protein-bacteria complex served as the imprint moiety. Atomic force microscopy and scanning electron microscopy were used to image the micron-size rod-shaped bacteria imprints formed on the silica surface. Fluorescence microscopy, which was used to follow the fabrication process with GFP as the representative protein, clearly demonstrated protein imprinting, protein removal and protein rebinding as well as protein specificity. Visible absorption spectroscopy using CYC as the imprint protein demonstrated relatively fast uptake kinetics and also good specificity against other proteins including bovine serum albumin (BSA), horseradish peroxidase (HRP), glucose oxidase (GOD), and lysozyme (LYZ). Collectively this work demonstrates a new surface bio-imprinting approach that generates recognition sites for proteins and provides a viable means to increase the binding capacity of such imprinted thin films.
机译：使用等级印记策略用于在具有高结合能力的硅酸盐膜中产生蛋白质印记以及对印记蛋白的选择性以及对其他蛋白质的特异性。在这项工作的第一部分中，使用棒状细菌作为模板，以在各种厚度的二氧化硅膜中产生印记，以打开二氧化硅框架并增加暴露于溶液的表面积。在第二部分中，蛋白质（例如，细胞色素C（Cyc）或绿色荧光蛋白（GFP））与枯草芽孢杆菌的表面共价附着在枯草芽孢杆菌的表面上，并且该蛋白质 - 细菌复合物用作压印部分。原子力显微镜和扫描电子显微镜用于在二氧化硅表面上形成形成的微米尺寸棒状细菌印记。荧光显微镜，其用于跟随具有GFP作为代表性蛋白的制造过程，清楚地证明了蛋白质印记，蛋白质去除和蛋白质refinding以及蛋白质特异性。使用Cyc作为压印蛋白质的可见吸收光谱表现出相对快速的吸收动力学以及对其他蛋白质的良好特异性，包括牛血清白蛋白（BSA），辣根过氧化物酶（HRP），葡萄糖氧化酶（GOD）和溶菌酶（Lyz）。本作集体证明了一种新的表面生物印迹方法，用于产生蛋白质的识别位点，并提供一种增加这种印迹薄膜的结合能力的可行方法。

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