We explore a bioinspired approach to design tailored functionalized capillary electrophoresis (CE) surfaces based on covalent grafting for biomolecules analysis. First, the approach aims to overcome well-known common obstacles in CE protein analysis affecting considerably the CE performance (asymmetry, resolution, and repeatability) such as the unspecific adsorption on fused silica surface and the lack of control of electroosmotic flow (EOF). Then, our approach, which relies on new amino-amide mimic hybrid precursors synthesized by silylation of amino-amides (Si–AA) derivatives with 3-isocyanatopropyltriethoxysilane, aims to recapitulate the diversity of protein–protein interactions (π–π stacking, ionic, Van der Waals…) found in physiological condition (bioinspired approach) to improve the performance of CE protein analysis (electrochromatography). As a proof of concept, these silylated Si–AA (tyrosinamide silylation, serinamide silylation, argininamide silylation, leucinamide silylation, and isoglutamine silylation acid) have been covalently grafted in physiological conditions in different amount on bare fused silica capillary giving rise to a biomimetic coating and allowing both the modulation of EOF and protein–surface interactions. The analytical performances of amino-amide functionalized capillaries were assessed using lysozyme, cytochrome C and ribonuclease A and compared to traditional capillary coatings poly(ethylene oxide), poly(diallyldimethylammonium chloride), and sodium poly(styrenesulfonate). EOF, protein adsorption rate, protein retention factor k, and selectivity were determined for each coating. All results obtained showed this approach allowed to modulate the EOF, reduce unspecific adsorption, and generate specific interactions with proteins by varying the nature and the amount of Si–AA in the functionalization mixture.

中文翻译：

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使用硅烷化氨基酰胺块和共价接枝调节毛细管电泳中电渗流和蛋白质-表面相互作用的仿生方法

我们探索一种基于生物分子分析的共价接枝设计定制的功能化毛细管电泳（CE）表面的仿生方法。首先，该方法旨在克服 CE 蛋白质分析中众所周知的常见障碍，这些障碍显着影响 CE 性能（不对称性、分​​辨率和重复性），例如熔融石英表面的非特异性吸附和电渗流 (EOF) 缺乏控制。然后，我们的方法依赖于通过氨基酰胺（Si-AA）衍生物与3-异氰酸基丙基三乙氧基硅烷的甲硅烷基化合成的新型氨基酰胺模拟杂化前体，旨在概括蛋白质-蛋白质相互作用的多样性（π-π堆积、离子、范德华……）在生理条件下（仿生方法）发现可以提高 CE 蛋白质分析（电色谱）的性能。作为概念证明，这些硅烷化的 Si-AA（酪氨酸酰胺硅烷化、丝氨酰胺硅烷化、精氨酰胺硅烷化、亮氨酰胺硅烷化和异谷氨酰胺硅烷化酸）已在生理条件下以不同的量共价接枝到裸露的熔融石英毛细管上，从而形成仿生涂层并允许调节 EOF 和蛋白质-表面相互作用。使用溶菌酶、细胞色素 C 和核糖核酸酶 A 评估氨基酰胺功能化毛细管的分析性能，并与传统毛细管涂层聚环氧乙烷、聚二烯丙基二甲基氯化铵和聚苯乙烯磺酸钠进行比较。确定每个涂层的EOF、蛋白质吸附率、蛋白质保留因子k和选择性。获得的所有结果表明，这种方法可以通过改变功能化混合物中 Si-AA 的性质和数量来调节 EOF，减少非特异性吸附，并与蛋白质产生特异性相互作用。