Select Bio EV-based Diagnostics, Delivery & Therapeutics 2020

EV-based Diagnostics, Delivery & Therapeutics 2020

 

Our 2020 presence at Select Bio’s EV-based Diagnostics, Delivery & Therapeutics will focus on standardization of EV characterization. Over 3 presentations, we will discuss the importance of single vesicle resolution in understanding EV heterogeneity. We will dive into recently published MISEV and MIFLOWCYT-EV guidelines which create a framework for generating and reporting data that will be interpretable across time and instrumentation. Finally, we will present a workshop with a live demonstration of our simple and reliable approach to standardized measurement of EV concentration, size, and molecular cargo, vFC™. In the workshop we will cover evaluation of the sensitivity of an instrument, discuss sample preparation, and describe analysis workflows highlighting the role of calibrators, standards, and controls in the workflow. See our schedule of events below. Click the links below to download the slides from our presentations.

Deciphering the Vesicle Code: Making Sense of EV Heterogeneity – 11:15-11:45 Monday

Cells release extracellular vesicles (EVs) that can carry molecular cargo, including lipids, proteins, and nucleic acids, to nearby or distant cells and affect their function. Understanding the mechanisms of EV biogenesis, uptake and transport has the potential to lead to new biomarkers, diagnostics, and therapeutics. A limit to realizing this potential is the ability to measure individual EVs and their cargo quantitatively and reproducibly. Conventional bulk biochemical analyses, which report only the total amount of cargo in an EV preparation, cannot effectively assess compositional heterogeneity. Single EV analysis methods are required, but conventional tools are challenged to accurately measure small, dim particles. We have developed a vesicle flow cytometry (vFC) method that can detect and size individual EVs to 70 nm, and measure surface cargo to ~25 molecules/EV. vFC allows the identification and characterization of vesicle sub-types within heterogeneous populations in culture supernatants of biofluids. Single EV analysis using vFC reveals striking differences in the expression of canonical vesicle cargo such as tetraspanins, as well as other surface markers including integrins, tumor markers, and carbohydrates on EVs in biofluids and even EVs collected from cultured cell lines. vFC-based single EV immunophenotyping will enable us to understand the origins and functions of EVs in much the same way cell-based immunophenotyping has enabled a detailed understanding of the immune system, and lead to a new generation of EV-based biomarkers, diagnostics, and therapeutics.

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Exosomes-EV Training Course 745pm Monday

The training course will provide an overview and discussion of various methods for the enrichment and characterization of exosomes, microvesicles, and other classes of extracellular vesicles (EVs). It will also provide an overview of various classes of circulating biomarkers carried by EVs.  Detailed protocols are presented to describe current approaches common to EV research. Comparison of existing technologies will focus on application of methods appropriately aligned to experimental questions. Experts in the field will lead Q&A at the conclusion of the program.

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vFC™ Workshop – 1-2pm Tuesday

Progress in EV research depends on methods to measure EVs and their cargo, but existing methods suffer from a lack of sensitivity, specificity and reproducibility. To help address this, ISEV (the International Society for Extracellular Vesicles) and the EV-FC working group have developed guidelines for EV analysis to promote rigor and reproducibility in EV research. In this Workshop, we will review the rational and goals of single EV characterization. The workshop will conclude with a live demonstration of a simple and reliable assay, Vesicle Flow Cytometry (vFC™), that enables sensitive and specific EV counting, sizing, and molecular cargo analysis. We will cover evaluation of the sensitivity of an instrument for EV analysis, discuss sample preparation and analysis workflows for vFC™, and highlight the role of calibrators, standards and controls in producing results that are reproducible over time and across instruments, and that are compliant with new guidelines and consensus best practices in the field.

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