Chromatographic Microfluidic Separation of Glycoprotein

描述（由申请人提供）：填充有自组装二氧化硅纳米颗粒的模制塑料微流体装置可以潜在地显著提高糖基化蛋白的梯度洗脱液相色谱分离的分辨率，同时消耗微量样品。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在2D凝胶。已经制造了与分离方法的所有常见的移动的相兼容的原理原型塑料微流体装置的证明。该装置潜在地允许自组装纳米颗粒在通道中作为色谱柱材料的优化填充。该提案项目的第一阶段侧重于开发填充通道的方法，以及二氧化硅颗粒的表面改性，以优化使用光学检测的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的纳米喷雾质谱仪，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.