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

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.