TOPIC 420 INTEGRATED PLATFORM FOR IN-DEPTH SINGLE-CELL PROTEOMICSPERIOD OF PERFORMANCE SEPTEMBER 13, 2021-JUNE 12, 2022FY21 PHASE I SBIR

TOPIC 420 深度单细胞蛋白质组学综合平台 性能周期 2021年9月13日-2022年6月12日 FY21 第一期 SBIR

• 批准号: 10610297
• 负责人: RYAN KELLY
• 金额: $ 5.5万
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-13 至 2022-06-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10610297
• 关键词: Automation; Cancer Diagnostics; Cells; Computer software; Custom; Development; Hela Cells; Impairment; Malignant Neoplasms; Manuals; Measurement; Neoplasm Circulating Cells; Performance; Phase; Positioning Attribute; Preparation; Proteins; Proteome; Recovery; Reproducibility; Robotics; Sampling; Small Business Innovation Research Grant; Structure; System; Tissues; anticancer research; biological systems; cost; data quality; instrument; liquid chromatography mass spectrometry; nanolitre; nanolitre scale; open source; protein expression; single cell proteins; targeted treatment; tumor heterogeneity

Proteins are key functional and structure components of biological systems including cancers, and are the primary targets of therapeutics. The ability to directly profile protein expression for cancer research and diagnostics has been impaired by the large amounts of tissue required to perform a measurement. These bulk analyses obscure important tumor heterogeneity and are unable to profile rare but important cells such as circulating tumor cells. We have recently developed a nanoliter-scale sample preparation platform termed nanoPOTS. When combined with ultrasensitive analysis, nanoPOTS enables profiling of hundreds to >1000 proteins from single cells but currently involves several manual steps and multiple custom instruments that make it unsuitable for broad dissemination. We will develop an integrated robotic platform that accomplishes both nanoliter-scale sample preparation and nanoliter-scale interfacing with LC/MS analysis. We will also evaluate alternative ‘nanowell’ substrate materials for nanoPOTS processing and a simplified one-step processing workflow. The system will be characterized in terms of achievable proteome coverage, reproducibility and throughput using single HeLa cells. Data quality will be assessed using open-source software. We expect to identify >800 proteins in single HeLa cells, positioning the system for full automation and increased measurement throughput during the subsequent phase of development.

蛋白质是包括癌症在内的生物系统的关键功能和结构组成部分，也是治疗的主要目标。用于癌症研究和诊断的直接分析蛋白质表达的能力已因执行测量分析所需的大量组织而受到损害。我们最近开发了一种称为 nanoPOTS 的纳升级样品制备平台，与超灵敏分析相结合，nanoPOTS 能够对数百至 > 1000 种蛋白质进行分析。我们将开发一个集成的机器人平台，该平台可完成纳升级样品制备和纳升级与 LC/MS 分析的连接。评估用于 nanoPOTS 处理的替代“纳米孔”基质材料和简化的一步处理工作流程 该系统将使用单个 HeLa 细胞在可实现的蛋白质组覆盖度、再现性和通量方面进行表征。我们期望在单个 HeLa 细胞中识别超过 800 种蛋白质，从而使系统在后续开发阶段实现完全自动化并提高测量通量。