SBIR PHASE II, TOPIC 420: INTEGRATED PLATFORM AND CONSUMABLES FOR ROBUST, SENSITIVE AND HIGH-THROUGHPUT SINGLE-CELL PROTEOMICS

基于质谱的蛋白质组学现在可以从包括单细胞在内的痕量样品中定量多达数千种蛋白质，但此类研究需要高度专业化的定制耗材和仪器，因此大多数实验室无法实现。现在存在商业解决方案，用于单细胞蛋白质组学（SCP）工作流程开始时的细胞分离和试剂分配，工作流程结束时可使用高灵敏度质谱，但中间存在很大差距 包括样品引入、分离和电离。我们的目标是为SCP和其他痕量生物分析的非专业最终用户提供强大的样品-答案工作流程。我们将开发（1）经过验证的样品处理试剂和耗材，用于简化样品制备;（2）专门用于SCP的高效液相色谱柱;（3）用于高通量分离的强大多柱LC平台。我们将实现每个细胞蛋白质组覆盖深度> 2，500个蛋白质， 每天96个细胞的通量（15分钟/细胞）。该平台还将与多路复用工作流程兼容，例如串联质量标签，以实现更高的吞吐量。因此，我们将建立一个创新平台，推进生物医学研究，生物标志物发现，诊断和个性化治疗。

Mass spectrometry-based proteomics can now quantify up to thousands of proteins from trace samples including single cells, yet such studies require highly specialized, custom-built consumables and instrumentation, and are thus out of reach of most laboratories. Commercial solutions now exist for cell isolation and reagent dispensing at the beginning of the single-cell proteomics (SCP) workflow, and highly sensitive mass spectrometry is available at the end of the workflow, but there is a large gap in the middle comprising sample introduction, separations and ionization. Our objective is to provide robust sample-to answer workflows for non-specialist end users for SCP and other trace bioanalyses. We will develop (1) validated sample processing reagents and consumables for streamlined sample preparation; (2) high performance liquid chromatography columns specifically for SCP; (3) a robust multi-column LC platform for high throughput separations. We will achieve a depth of proteome coverage >2,500 proteins per cell and a throughput of 96 cells per day (15 min/cell). The platform will also be compatible with multiplexed workflows such as tandem mass tags for an order of magnitude higher throughput. We will thus establish an innovative platform for advancing biomedical research, biomarker discovery, diagnostics and individualized therapy.