High sensitivity reagents for bioassays employing mass cytometry

点击翻译按钮获取中文摘要

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.