Chromatographic Microfluidic Separation of Glycoprotein

描述(由申请人提供)：填充自组装二氧化硅纳米颗粒的模塑塑料微流控设备可以极大地提高糖基化蛋白质的梯度洗脱液相分离的分辨率，同时消耗极少量的样品。在同一设备中使用多个通道(超过100个)将极大地提高吞吐量。这种分离补充了现有的使用等电聚焦和2D凝胶中的分子筛分的二维蛋白质分离。制作了与各种常见流动相分离方法兼容的塑料微流控器件的原理验证样机。该设备潜在地允许在通道中优化填充自组装纳米颗粒作为色谱柱材料。该提案项目的第一阶段重点是开发填充通道的方法，并对二氧化硅颗粒进行表面修饰，以使用光学检测优化模型糖基化蛋白质的分离。在该项目的第二阶段，将开发具有多个分离通道接口的纳米质谱分析设备，以提高吞吐量和检测灵敏度。

DESCRIPTION (provided by applicant): Molded plastic microfluidic devices packed with self-assembled silica nanoparticles can potentially dramatically improve the resolution of the gradient elution liquid chromatographic separation of glycosylated proteins while consuming minute amount of sample. Using multiple channels (over 100) in the same device will greatly enhance throughput. This separation complements existing two-dimensional protein separations using isoelectric focusing and molecular sieving in a 2D gel. A proof of principle prototype plastic microfluidic device that is compatible with all common mobile phases of the separation methods has been fabricated. This device potentially allows the optimized packing of self-assembled nanoparticles in the channel as chromatographic column material. The Phase 1 of this proposal project focuses on developing methods for packing the channels, and surface modification of the silica particles to optimize the separation of model glycolsylated proteins using optical detection. In Phase II of the project, devices with multiple separation channels interfaced for nanospray mass spectrometry will be developed to increase throughput and detection sensitivity.