Lee K, Itharaju R R, Puleo D A
Center for Biomedical Engineering, University of Kentucky, Lexington, KY, USA.
Acta Biomater. 2007 Jul;3(4):515-22. doi: 10.1016/j.actbio.2007.01.003. Epub 2007 Mar 23.
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 0.3microgm(-2) for low loading of RNase to 152microgm(-2) for 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.
分子印迹是一种用于在材料表面创建特定识别位点的技术。尽管该技术已广泛应用于小分子的色谱分离，但在生物材料应用方面尚未得到充分研究。因此，这些实验的目的是探索分子印迹技术在制备优先结合特定蛋白质的生物材料方面的潜力。使用溶胶-凝胶工艺，用溶菌酶或核糖核酸酶A对大孔聚硅氧烷（二氧化硅）支架进行印迹。通过改变溶胶中加载的蛋白质量来改变与潜在结合位点数相关的表面可及蛋白量。高达62%的加载蛋白是可及的。每单位表面积的蛋白量范围从低加载量核糖核酸酶时的0.3μg/m²到高加载量溶菌酶时的152μg/m²。然后，当在同时含有印迹蛋白和大小相当（约14kD）的竞争蛋白的混合物中孵育时，评估蛋白印迹支架优先识别模板生物分子的能力。在含有单一蛋白的溶液中，与竞争蛋白相比，结合的模板最多多3.6倍。此外，在含有等量两种分子的溶液中，多孔支架结合的模板比竞争蛋白多多达三倍，这一优先结合水平与分子印迹文献中报道的有机和无机材料的值相似。
