Protein-imprinted polysiloxane scaffolds.
Protein-imprinted polysiloxane scaffolds.
复制标题
DOI：
10.1016/j.actbio.2007.01.003
复制
发表时间：
2007-07
影响因子：
9.7
通讯作者：
K. Lee;R. R. Itharaju-R.;D. Puleo
中科院分区：
文献类型：
--
作者：
K. Lee;R. R. Itharaju-R.;D. Puleo
分子印迹是一种用于在材料表面产生特异性识别位点的技术。虽然广泛开发的色谱分离的小分子，这种方法还没有充分研究生物材料的应用。因此，这些实验的目的是探索分子印迹用于创建优先结合特定蛋白质的生物材料的潜力。大孔聚硅氧烷（二氧化硅）支架使用溶胶-凝胶工艺与溶菌酶或RNA酶A印迹。表面可及蛋白质的量，这是有关的潜在结合位点的数量，通过改变加载到溶胶中的蛋白质的量而变化。高达62%的负载蛋白是可获得的。每单位表面积的蛋白质量范围从0.3μgm− 2（低载量RNA酶）到152μgm− 2（高载量溶菌酶）。然后评估蛋白质印迹支架在含有印迹蛋白质和相当大小（约14 kD）的竞争蛋白质的混合物中孵育时优先识别模板生物分子的能力。在含有单一蛋白质的溶液中，与竞争对手相比，结合的模板高达3.6倍。此外，在含有等量的两种分子的溶液中，多孔支架结合的模板比竞争蛋白多三倍，这是一种与有机和无机材料的分子印迹文献中报道的值相似的优先结合水平。
Molecular imprinting is a technique used to create specific recognition sites on the surface of materials. Although widely developed for chromatographic separation of small molecules, this approach has not been adequately investigated for biomaterial applications. Thus, the objective of these experiments was to explore the potential of molecular imprinting for creating biomaterials that preferentially bind specific proteins. Macroporous polysiloxane (silica) scaffolds were imprinted with either lysozyme or RNase A using sol–gel processing. The quantity of surface-accessible protein, which was related to the number of potential binding sites, was varied by changing the amount of protein loaded into the sol. Up to 62% of loaded protein was accessible. The amount of protein per unit surface area ranged from 0.3μgm−2for low loading of RNase to 152μgm−2for high loading of lysozyme. Protein-imprinted scaffolds were then evaluated for their ability to preferentially recognize the template biomolecule when incubated in mixtures containing both the imprinted protein and a competitor protein of comparable size (approximately 14kD). In solutions containing a single protein, up to 3.6 times more template bound compared with the competitor. Furthermore, in solutions containing equal amounts of both molecules, the porous scaffolds bound up to three times more template than the competitor protein, which is a level of preferential binding similar to values reported in the molecular imprinting literature for both organic and inorganic materials.