Silica Colloidal Crystals for High Resolution MALDI-MS of Glycoproteins

描述(申请人提供)：该产品是一种涂有二氧化硅胶体晶体的玻片，将蛋白质电泳与基质辅助激光解吸/电离质谱仪(MALDI-MS)检测结合在一起，在两者中都具有优越的性能。第二阶段的建议是开发一种仪器，用于在Intat糖蛋白的蛋白质组学中方便地应用载玻片。这项提议是普渡大学的Wirth小组和Li-COR生物科学公司之间的合作，前者开发了可与MALDI-MS结合的蛋白质分离新介质，后者的核心能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衬底的初步结果显示，加合物的形成减少，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媒介。结合使用能够实现高分辨率蛋白质分离的单一薄层晶体表面和改进的MALDI-MS检测来分离和检测蛋白质的能力，将扩大使用MALDI-MS识别基于翻译后修饰的新生物标记物的用途。第一阶段的具体目标是优化二氧化硅晶体的性质，表征适用的相对分子质量范围，研究使用这种底物提高分辨率和减少加合物形成的直接结果，并将蛋白质分离技术与MALDI-MS检测结合在单个二氧化硅胶体晶体表面上。

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.