Silica Colloidal Crystals for High Resolution MALDI-MS of Glycoproteins

描述（由申请人提供）：该产品是涂有二氧化硅胶体晶体的载玻片，用于结合蛋白质电泳和基质辅助激光解吸/电离质谱（MALDI-MS）检测，两者都具有优异的性能。II期计划将开发一种仪器，方便地将载玻片应用于体内糖蛋白的蛋白质组学。该提案是普渡大学的Wirth小组和LI-COR生物科学公司之间的合作，前者开发了用于蛋白质分离的新媒体，可以与MALDI-MS结合，后者的核心竞争力是生物分析仪器的商业化。使用改性二氧化硅胶体晶体表面作为MALDI-MS底物的初步结果表明，加合物形成减少，质量分辨率提高。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。结合使用单薄层晶体表面分离和检测蛋白质的能力，实现高分辨率蛋白质分离和改进的MALDI-MS检测，将扩大使用MALDI-MS识别基于翻译后修饰的新型生物标志物的实用性。第一阶段的具体目标是优化二氧化硅晶体性能，表征适用的分子量范围，研究使用该底物的直接结果是分辨率的提高和加合物形成的减少，并将蛋白质分离技术与MALDI-MS检测在单个二氧化硅胶体晶体表面结合起来。

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.