Protein analysis by desorption electrospray ionization mass spectrometry

通过解吸电喷雾电离质谱法进行蛋白质分析

• 批准号: 1508626
• 负责人: Andre Venter
• 金额: $ 29.86万
• 依托单位: Western Michigan University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1508626&HistoricalAwards=false
• 关键词: Protein; analysis; desorption; electrospray; ionization

With this award, the Chemical Measurement and Imaging Program in the Division of Chemistry is funding Professor Andre Venter at Western Michigan University to investigate various methods to improve the analysis of protein molecules by desorption electrospray ionization mass spectrometry (DESI-MS). Imaging of tissue samples by DESI is an important new tool to aid in biological tissue imaging. DESI-imaging applications are currently limited to the analysis of small molecules such as lipids, fatty acids, and small molecules. Developing the ability to analyze proteins would extend the capabilities of imaging DESI-MS to determine the spatial distribution of proteins in biological relevant tissues. In addition, Professor Venter will make significant efforts to increase participation of undergraduate students, especially from underrepresented groups in research, and educate the general public about the importance of chemistry in their everyday life through collaboration with the Gwen Frostic School of Art at WMU.Currently when proteins are analyzed by DESI-MS, sensitivity decreases with increasing molecular weight, and poor protein dissolution has been implicated as the major cause. Methods to improve dissolution and desorption such as the application of heat, the addition of a variety of additives to the DESI spray and a delayed desorption DESI sprayer will be investigated separately and in combination. To decipher mechanistic aspects of these mitigation strategies, Professor Venter will separately investigate the effects on dissolution of each, during desorption and ionization by using their previously developed Spray Desorption Collection, and Reflective Electrospray Ionization methods. Construction and characterization of a dual-sprayer DESI configuration will be carried out to determine optimum delay between localized surface wetting and desorption to maximize signal to noise ratios of proteins during DESI analysis, while conserving lateral resolution.

化学系的化学测量和成像项目获得这一奖项，资助西密歇根大学的安德烈·文特尔教授研究各种方法，以改进解吸电喷雾电离质谱仪(DESI-MS)对蛋白质分子的分析。利用DESI对组织样品进行成像是辅助生物组织成像的重要新工具。目前，Desi成像应用仅限于分析小分子，如脂类、脂肪酸和小分子。发展分析蛋白质的能力将扩展成像Desi-MS的能力，以确定生物相关组织中蛋白质的空间分布。此外，文特尔教授将做出重大努力，增加本科生，特别是来自未被充分代表的群体参与研究，并通过与威斯康星大学格温·弗罗斯特艺术学院的合作，教育普通公众化学在他们日常生活中的重要性。目前，当用DESI-MS分析蛋白质时，灵敏度随着分子量的增加而降低，蛋白质溶解不良被认为是主要原因。改善溶解和解吸的方法，如加热、在DESI喷雾器中添加各种添加剂和延迟解吸Desi喷雾器将单独和联合进行研究。为了破译这些缓解策略的机制方面，文特尔教授将使用他们之前开发的喷雾解吸收集和反射式电喷雾电离方法，分别研究解吸和电离过程中每种策略对溶解的影响。将进行双喷雾器DESI配置的构建和表征，以确定局部表面润湿和解吸之间的最佳延迟，以最大限度地提高DESI分析期间蛋白质的信噪比，同时保持横向分辨率。