Silica Colloidal Crystals for High Resolution MALDI-MS of Glycoproteins

描述（由申请方提供）：本品是一种涂有二氧化硅胶体晶体的载玻片，可将联合收割机蛋白质电泳与基质辅助激光解吸/电离质谱（MALDI-MS）检测结合起来，在两者中均具有上级性能。第二阶段的建议将是开发一种仪器，方便应用的幻灯片在蛋白质组学的intat糖蛋白。该提案是普渡大学Wirth小组与LI-COR，Biosciences的合作，前者开发了可与MALDI-MS结合的蛋白质分离新介质，后者的核心竞争力是生物分析仪器的商业化。使用改性的二氧化硅胶体晶体表面作为MALDI-MS底物的初步结果表明，减少加合物的形成，并增加质量分辨率。此外，二氧化硅胶体晶体表面是使用等电聚焦分离蛋白质的介质。结合使用能够实现高分辨率蛋白质分离的单个薄层结晶表面分离和检测蛋白质的能力和改进的MALDI-MS检测将扩展使用MALDI-MS用于基于翻译后修饰鉴定新生物标志物的效用。第一阶段的具体目标是优化二氧化硅晶体的性质，表征适用的分子量范围，研究作为使用该底物的直接结果的提高的分辨率和加合物形成的减少，以及将蛋白质分离技术与单个二氧化硅胶体晶体表面上的MALDI-MS检测联合收割机相结合。

DESCRIPTION (provided by applicant): The product is a slide coated with silica colloidal crystals to combine protein electrophoresis with matrix- assisted laser desorption/ionization mass spectrometry (MALDI-MS) detection, giving superior performance in both. The Phase II proposal will be to develop an instrument for facile application of the slide in proteomics of intat glycoproteins. This proposal is a collaboration between the Wirth group at Purdue, who have developed new media for protein separations that can be coupled with MALDI-MS, and LI-COR, Biosciences, whose core competency is the commercialization of instruments for bioanalysis. Preliminary results using a modified silica colloidal crystalline surface as a MALDI-MS substrate show a reduction in adduct formation, and an increase in mass resolution. Additionally, the silica colloidal crystalline surface is a medium to separate proteins using isoelectric focusing. Combining the ability to both separate and detect proteins using a single thin layer crystalline surface that enables high resolution protein separation and improved MALDI-MS detection will expand the utility of using MALDI-MS for identifying novel biomarkers based on post translational modifications. The phase I specific aims are to optimize the silica crystal propertie, to characterize the applicable molecular weight range, to study the enhanced resolution and the reduction in adduct formation as a direct result of using this substrate, and to combine protein separation techniques with MALDI-MS detection on a single silica colloidal crystalline surface.