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首页> 外文期刊>ACS applied materials & interfaces >Pore-Size Dependent Protein Adsorption and Protection from Proteolytic Hydrolysis in Tailored Mesoporous Silica Particles
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Pore-Size Dependent Protein Adsorption and Protection from Proteolytic Hydrolysis in Tailored Mesoporous Silica Particles

机译：量身定制的介孔二氧化硅颗粒中孔径依赖蛋白的吸附和蛋白水解保护。

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摘要

Protein adsorption and interactions with mesoporous silica are of interest for a broad range of applications including drug delivery, chemical synthesis, biosensors, and bioseparations. A major challenge in designing mesoporous silica supports for tailored protein interaction is the differentiation of protein interactions at the surface of the particle from interactions within the pore, important features when considering mesoporous silica as a protective support for active proteins. In this investigation, the location of Enhanced Green Fluorescent Proteins (EGFPs) adsorbed on tailored mesoporous silica particles is examined as a function of pore diameter using proteolytic hydrolysis to distinguish between accessible and inaccessible proteins. Pore size control is achieved by tuning the hydrothermal aging temperature (60—110 °C) during synthesis, where the synthesis results in 5-15 μm diameter spherical particles appropriate for imaging by confocal scanning laser microscopy (CSLM). In low pH environments, EGFP unfolds within pores and on the surface of particles, rendering it susceptible to proteolytic hydrolysis by the protease Pepsin A. Upon return to neutral pH, un-hydrolyzed EGFP regains its fluorescence and can be visualized within the mesoporous particles. The pore-size dependent loading and protection of EGFP (2.4 nm diameter X 4.2 nm) from proteolytic attack by Pepsin A (7.3 nm X 3.6 nm X S.4 nm) is demonstrated by the retention of fluorescence in 7.3 nm pores. Larger-pored materials (> 9 nm) provide diminishing protection for EGFP, and the protection is greatly reduced with increasing pore size and pore size distribution breadth. Proteolytic hydrolysis is used to delineate the activity of pore-loaded versus surface-bound proteins and to establish that there is an optimal pore diameter for loading EGFP while protecting it from attack by a larger proteolytic enzyme.
机译：蛋白质吸附和与中孔二氧化硅的相互作用对于包括药物递送，化学合成，生物传感器和生物分离在内的广泛应用是令人感兴趣的。设计用于定制蛋白质相互作用的介孔二氧化硅载体的主要挑战是区分颗粒表面的蛋白质相互作用与孔内的相互作用，当将介孔二氧化硅用作活性蛋白质的保护性载体时，这是重要的特征。在这项研究中，通过蛋白水解水解来区分可及和不可及蛋白质，研究了吸附在特制介孔二氧化硅颗粒上的增强型绿色荧光蛋白质（EGFP）的位置与孔径的关系。通过调节合成过程中的水热时效温度（60-110°C），可以控制孔径，合成过程中会产生直径5-15μm的球形颗粒，适合通过共聚焦扫描激光显微镜（CSLM）进行成像。在低pH的环境中，EGFP在孔中和颗粒表面上展开，使其易于被蛋白酶胃蛋白酶A进行蛋白水解。回到中性pH值后，未水解的EGFP恢复其荧光并可以在中孔颗粒中看到。通过荧光在7.3 nm孔中的保留，可以证明EGFP（2.4 nm直径X 4.2 nm）的孔径依赖性加载和保护不受胃蛋白酶A（7.3 nm X 3.6 nm X S.4 nm）的蛋白水解攻击。较大孔径的材料（> 9 nm）对EGFP的保护作用减弱，并且随着孔径和孔径分布宽度的增加，保护作用大大降低。蛋白水解用于描述孔加载蛋白和表面结合蛋白的活性，并确定在加载EGFP时有一个最佳孔径，同时保护它不受较大蛋白水解酶的攻击。

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