Chromatographic Microfluidic Separation of Glycoprotein

描述（由申请人提供）：包装有自组装二氧化硅纳米颗粒的模压塑料微流控装置可以在消耗微量样品的情况下显著提高梯度洗脱液相色谱分离糖基化蛋白的分辨率。在同一设备中使用多个通道（超过100个）将大大提高吞吐量。这种分离补充了现有的二维蛋白质分离使用等电聚焦和分子筛选在二维凝胶。制作了一个原理验证原型塑料微流控装置，该装置与所有常用的分离方法的流动相兼容。该装置有可能使自组装纳米颗粒在通道中作为色谱柱材料的最佳填料。本项目第一阶段的重点是开发通道包装方法，以及二氧化硅颗粒的表面修饰，以优化使用光学检测的模型糖基化蛋白的分离。在该项目的第二期，将开发具有多个分离通道接口的纳米喷雾质谱设备，以提高通量和检测灵敏度。

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.