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

• 批准号: 8781037
• 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
• 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/8781037
• 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; 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; public health relevance; software systems

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单一二氧化硅胶体晶体表面上的 MALDI-MS 检测相结合。