High sensitivity reagents for bioassays employing mass cytometry

用于采用质谱流式细胞仪进行生物测定的高灵敏度试剂

• 批准号: 484832-2015
• 负责人: Winnik, Mitchell
• 金额: $ 10.02万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Collaborative Research and Development Grants
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=693499
• 关键词: sensitivity; reagents; bioassays; employing; mass

This proposal describes the synthesis, characterization, and biofunctionalization of inorganic nanoparticles designed as high-sensitivity reagents for mass cytometry (CyTOF), a technique invented and commercialized by our partner Fluidigm Canada in Markham ON. Mass cytometry works on the principle of atomic mass spectrometry and employs reagents labeled with metal isotopes to detect proteins and other biomarkers expressed in or on individual cells. It enables one to examine individual cells at high throughput (up to 1000 cells/sec), and its greatest strength is that it can simultaneously measure many more biomarkers per cell than flow cytometry, which employs fluorescent reagents. A current shortcoming of mass cytometry is its limited sensitivity in detecting biomarkers present at a low copy number per cell. The research proposed here will overcome this limitation by developing nanoparticle reagents that carry many more copies of a metal isotope than the current reagents. We anticipate that nanoparticle reagents will provide one to two orders of magnitude increase in sensitivity. A major challenge in this research will be to modify the surface of these nanoparticles so that they only bind to targeted cells and have negligible non-specific interaction with non-targeted cells. In addition, we will develop nanoparticle reagents that will increase the number of metal isotopes that can be employed in an analysis and thus increase the number of different biomarkers that can be measured for each cell. We hope that these reagents in combination with mass cytometry will provide technology that one can use to understand, diagnose, and, hopefully, cure complex human diseases such as cancer, metabolic disorders, and heart disease.

该提案描述了无机纳米颗粒的合成、表征和生物功能化，该纳米颗粒被设计为质谱流式分析 (CyTOF) 的高灵敏度试剂，该技术由我们位于安大略省万锦市的合作伙伴 Fluidigm Canada 发明并商业化。质谱流式分析仪的工作原理是原子质谱分析，并采用金属同位素标记的试剂来检测单个细胞内或细胞上表达的蛋白质和其他生物标志物。它使人们能够以高通量（高达 1000 个细胞/秒）检查单个细胞，其最大优势在于它可以比使用荧光试剂的流式细胞术同时测量每个细胞更多的生物标志物。目前质谱流式细胞术的一个缺点是其在检测每个细胞中低拷贝数的生物标志物方面的灵敏度有限。这里提出的研究将通过开发比现有试剂携带更多金属同位素副本的纳米颗粒试剂来克服这一限制。我们预计纳米颗粒试剂将使灵敏度提高一到两个数量级。这项研究的一个主要挑战是修改这些纳米粒子的表面，使它们只与目标细胞结合，并且与非目标细胞的非特异性相互作用可以忽略不计。此外，我们将开发纳米颗粒试剂，增加可用于分析的金属同位素的数量，从而增加每个细胞可测量的不同生物标志物的数量。我们希望这些试剂与质谱流式细胞术相结合，能够提供一种技术，可用于理解、诊断，并有望治愈复杂的人类疾病，如癌症、代谢紊乱和心脏病。