A Novel Mass Spectrometric Ion Source for Proteomics

描述(申请人提供)：本研究的总体目标是：(1)开发一种新的高效液相色谱-电喷雾/基质辅助激光解吸/电离-质谱仪(LC-ESI/MALDI-MS)接口，以增强复杂混合物中蛋白质的质谱学鉴定和序列分析；(2)应用该技术解决生物研究前沿的问题。其具体目标是：(1)开发一种新的电离方法用于质谱学，该方法能够增加蛋白质组的覆盖率，同时为复杂混合物中的蛋白质的分析提供一种更快速和更廉价的方法，以及(2)应用所开发的技术来识别作为急性髓系白血病的诊断标记或潜在药物靶标的蛋白质。通过在大气压下操作，该技术将消除将MALDI与微毛细管高效液相色谱结合时遇到的许多障碍。该方法将利用ESI气溶胶液滴作为MALDI离子产生的平台。与其他任何高效液相色谱-质谱仪方法不同的是，这种方法将同时利用两种电离方法的优点。这项技术将被应用于确定与混合血统白血病基因(MLL)有关的调控途径，MLL是染色体易位的常见靶点，经常导致急性髓系白血病。利用HPLC-ESI/MALDI-MS，我们建议鉴定MLL-ENL融合蛋白诱导的蛋白质，这些蛋白质可以作为治疗急性髓系白血病的生物标志物或潜在的药物靶点。

DESCRIPTION (provided by applicant): The overall goals of this research are: (1) to develop a novel high performance liquid chromatography electrospray/ matrix-assisted laser desorption/ionization-mass spectrometry (HPLC-ESI/MALDI-MS) interface to enhance the mass spectrometric identification and sequence analysis of proteins in complex mixtures and (2) to employ this technology to solve problems at the forefront of biological research. The specific aims are to: (1) develop a novel ionization method for mass spectrometry that is capable of increasing proteome coverage, while at the same time providing a more rapid and inexpensive approach for the analysis of proteins in complex mixtures and (2) to apply developed technology to identify proteins that function as diagnostic markers or potential drug targets for acute myeloid leukemias. By operating at atmospheric pressure, the technique will eliminate many of the barriers encountered when combining MALDI with microcapillary HPLC. The method will utilize ESI aerosol droplets to serve as a platform for MALDI ion generation. And unlike any other HPLC-MS approach, this method will capitalize the favorable attributes of both ionization methods simultaneously. This technology will be applied to determine the regulatory pathways involved with the mixed lineage leukemia gene (MLL), which is a common target for chromosome translocation that often results in acute myeloid leukemias. Utilizing HPLC-ESI/MALDI-MS, we propose to identify proteins, induced by the MLL-ENL fusion protein that could function as biomarkers or potential drug targets for treatment of acute myeloid leukemias.