Silica Colloidal Crystals for High Resolution MALDI-MS of Glycoproteins

用于糖蛋白高分辨率 MALDI-MS 的二氧化硅胶体晶体

基本信息

批准号：
8927044
负责人：
Jon P Anderson
金额：
$ 30.65万
依托单位：
LI-COR BIOSCIENCES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8927044
关键词：
AFP gene Appearance Biological Markers Biological Sciences Biotechnology Caliber Characteristics Collaborations Colloids Coupled Crystallization Detection Development Diagnosis Dimensions Electrophoresis Glass Glycoproteins Goals Health In Situ Isoelectric Focusing Isoelectric Point Lasers Malignant Neoplasms Measures Methods Modification Molecular Weight Noise Normal Cell Peptides Performance Phase Post Translational Modification Analysis Post-Translational Protein Processing Primary carcinoma of the liver cells Proteins Proteomics Reagent Reproducibility Research Resolution Resources Sales Sampling Scanning Series Signal Transduction Silicon Dioxide Slide Specificity Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Surface System Techniques Technology Thick Work adduct alpha-Fetoproteins base cancer cell cancer risk commercialization design drug discovery improved instrument ionization miniaturize new technology novel protein complex software systems

项目摘要

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.

描述（申请人提供）：该产品是一种涂有二氧化硅胶体晶体的载玻片，将蛋白质电泳与基质辅助激光解吸/电离质谱（MALDI-MS）检测相结合，在两者方面均具有优异的性能。第二阶段的提案将是开发一种仪器，以便在完整糖蛋白的蛋白质组学中轻松应用载玻片。该提案是普渡大学 Wirth 团队和 LI-COR Biosciences 之间的合作，该团队开发了可与 MALDI-MS 结合的蛋白质分离新介质，LI-COR Biosciences 的核心能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。将使用单个薄层晶体表面分离和检测蛋白质的能力相结合，实现高分辨率蛋白质分离和改进的 MALDI-MS 检测，将扩展使用 MALDI-MS 识别基于翻译后修饰的新型生物标志物的效用。第一阶段的具体目标是优化二氧化硅晶体特性，表征适用的分子量范围，研究使用该底物直接导致的分辨率提高和加合物形成减少，并将蛋白质分离技术与 MALDI 相结合。单一二氧化硅胶体结晶表面的 MS 检测。