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：
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
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/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

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（由申请人提供）：该产品是带有二氧化硅胶体晶体的载体，可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，在两者中都具有出色的性能。第二阶段的提案将是开发一种用于在Intat糖蛋白蛋白质组学中易于应用的工具。该提案是普渡大学的Wirth集团之间的合作，他们开发了可以与Maldi-MS结合的蛋白质分离的新媒体，Li-Cor，Biosciences，其核心竞争力是对生物分析工具的商业化。使用改良的二氧化硅胶体晶体表面作为MALDI-MS底物的初步结果显示，加合物的形成减少，质量分辨率增加。另外，二氧化硅胶体晶体表面是使用等电聚焦分离蛋白质的培养基。使用单个薄层晶体表面结合分离和检测蛋白质的能力，从而实现高分辨率蛋白质分离和改进的MALDI-MS检测能力将扩大使用MALDI-MS根据翻译后修饰来识别新型生物标志物的实用性。第一阶段的特定目的是优化二氧化硅晶体性能，以表征适用的分子量范围，以研究增强的分辨率和加合物形成的减少，这是使用该底物的直接结果，并将蛋白质分离技术与单硅胶体表面上的MALDI-MS检测结合在一起。