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首页> 外文期刊>Chemistry: A European journal >Assembly-controlled biocompatible interface on a microchip: Strategy to highly efficient proteolysis
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Assembly-controlled biocompatible interface on a microchip: Strategy to highly efficient proteolysis

机译：芯片上由装配控制的生物相容性界面：高效蛋白水解的策略

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摘要

A biocompatible interface was constructed on a microchip by using the layer-by-layer (LBL) assembly of charged polysaccharides incorporating proteases for highly efficient proteolysis. The controlled assembly of natural polyelectrolytes and the enzyme-adsorption step were monitored by using a quartz-crystal microbalance and atomic force microscopy (AFM). Such a multilayer-assembled membrane provides a biocompatible interconnected network with high enzyme-loading capacity. The maximum digestion rate of the adsorbed trypsin in a microchannel was significantly accelerated to 1600 mm min(-1) mu g(-1), compared with the tryptic digestion in solution. Based on the Langmuir isotherm model, the thermodynamic constant of adsorption K was calculated to be 1.6 x 10(5) M-1 and the maximum adsorption loading Gamma(max) was 3.6 x 10(-6) mol m(-2), 30 times more than a monolayer of trypsin on the native surface. The tunable interface containing trypsin was employed to construct a microchip reactor for digestion of femtomoles of proteins and the produced peptides were analyzed by MALDI-TOF mass spectroscopy. The efficient on-chip proteolysis was obtained within a few seconds, and the identification of biological samples was feasible.
机译：通过使用带电荷的多糖的逐层（LBL）组装并结合了蛋白酶来进行高效蛋白水解，可在微芯片上构建生物相容性界面。通过使用石英晶体微量天平和原子力显微镜（AFM）监测天然聚电解质的受控组装和酶吸附步骤。这种多层组装的膜提供了具有高酶负载能力的生物相容性互连网络。与溶液中的胰蛋白酶消化相比，微通道中吸附的胰蛋白酶的最大消化速率显着提高到1600 mm min（-1）μg（-1）。根据Langmuir等温模型，吸附K的热力学常数经计算为1.6 x 10（5）M-1，最大吸附负荷Gamma（max）为3.6 x 10（-6）mol m（-2），天然表面上单层胰蛋白酶的30倍以上。包含胰蛋白酶的可调界面被用于构建微芯片反应器，用于消化蛋白质飞克分子，并通过MALDI-TOF质谱分析产生的肽。在几秒钟内获得了有效的芯片上蛋白水解，生物样品的鉴定是可行的。

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