No need for beads! Virus-derived biological standards for nanoscale flow cytometry

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Nanoscale flow cytometry (NFC) is an emerging and transformative technology that is designed to analyze the composition of cell-derived surface markers expressed on minute biological particles, most notably viruses and extracellular vesicles (EVs). EVs resemble tiny bubbles constantly released by all cell types and serve as a means of communication to coordinate growth, differentiation, but also to warn other cells of potential dangers such as injury, infection and diseases such as cancer, Parkinson's and Alzheimer's. Analyzing the surface markers of EVs isolated from biological fluids such as blood, saliva and urine may provide early warning signs of disease or infection. As such, markers on the surface of EVs are poised to become valuable and powerful tools for medical and veterinary clinical diagnostics. Despite NFC being capable of analyzing EV surface markers, its broad deployment and adoption by hospitals and research centers has been hindered by the absence of suitable commercial instrument calibration standards and experimental controls. We have developed fluorescent virus-derived particles that mimic the size and physical properties of EVs, and thereby act as extremely reliable and effective controls for NFC. The viruses can be produced consistently, cheaply and in large quantities. The work proposed here will further improve these viruses to meet specific research and clinical requirements. The anticipated outcome is the production of 10 different virus-derived standards or controls that can be used in various research and clinical NFC applications for the accurate analysis of EV surface markers. We will ensure the supply of theses virus-derived particles through the creation of a start-up company dedicated to making them. The start-up will supply NFC instrument manufacturers and reagent distributors with the virus-derived standards, and they will make them available to their academic and industry clients. Economic and societal benefits to Canada include the training of highly qualified personnel, and the creation of a start-up that will manufacture a reagent for NFC applications which will, in turn, lead to a new generation of non-invasive diagnostic tests for disease and infection for use in humans, fish and livestock.**************