Bacteria assisted protein imprinting in sol–gel derived films,Analyst

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Bacteria assisted protein imprinting in sol–gel derived films
Analyst ( IF 3.6 ) Pub Date : 2017-12-20 00:00:00 , DOI: 10.1039/c7an01509g
Wei Cai _{1,

2,

3,

4} , Hui-Hui Li _{1,

2,

3,

4} , Zhe-Xue Lu _{1,

2,

3,

4} , Maryanne M. Collinson _{5,

6,

7,

8}

Affiliation

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.

中文翻译：

细菌辅助蛋白质在溶胶-凝胶衍生膜中的印迹

分级印迹策略用于在硅酸盐薄膜中创建蛋白质印迹，该印迹具有高的结合能力以及对印迹蛋白质的选择性，而对其他蛋白质的特异性却很小。在这项工作的第一部分中，棒状细菌被用作模板，以在各种厚度的二氧化硅膜上形成印记，以打开二氧化硅骨架并增加暴露于溶液中的表面积。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））共价附着于枯草芽孢杆菌的表面并且该蛋白质-细菌复合物用作印迹部分。原子力显微镜和扫描电子显微镜用于成像在二氧化硅表面形成的微米级棒状细菌印记。荧光显微镜用于跟踪以GFP为代表蛋白的制备过程，清楚地证明了蛋白印迹，蛋白去除和蛋白重新结合以及蛋白特异性。使用CYC作为印迹蛋白的可见吸收光谱显示出相对较快的吸收动力学，并且对其他蛋白（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））具有良好的特异性。

更新日期：2017-12-20

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