Critical considerations in determining the surface charge of small extracellular vesicles

Author:

Tamrin Sara Hassanpour123ORCID,Phelps Jolene12ORCID,Nezhad Amir Sanati23ORCID,Sen Arindom12ORCID

Affiliation:

1. Pharmaceutical Production Research Facility, Department of Chemical and Petroleum Engineering, Schulich School of Engineering University of Calgary Calgary Alberta Canada

2. Department of Biomedical Engineering, Schulich School of Engineering University of Calgary Calgary Alberta Canada

3. BioMEMS and Bioinspired Microfluidic Laboratory, Department of Biomedical Engineering, Schulich School of Engineering University of Calgary Calgary Alberta Canada

Abstract

AbstractSmall extracellular vesicles (EVs) have emerged as a focal point of EV research due to their significant role in a wide range of physiological and pathological processes within living systems. However, uncertainties about the nature of these vesicles have added considerable complexity to the already difficult task of developing EV‐based diagnostics and therapeutics. Whereas small EVs have been shown to be negatively charged, their surface charge has not yet been properly quantified. This gap in knowledge has made it challenging to fully understand the nature of these particles and the way they interact with one another, and with other biological structures like cells. Most published studies have evaluated EV charge by focusing on zeta potential calculated using classical theoretical approaches. However, these approaches tend to underestimate zeta potential at the nanoscale. Moreover, zeta potential alone cannot provide a complete picture of the electrical properties of small EVs since it ignores the effect of ions that bind tightly to the surface of these particles. The absence of validated methods to accurately estimate the actual surface charge (electrical valence) and determine the zeta potential of EVs is a significant knowledge gap, as it limits the development of effective label‐free methods for EV isolation and detection. In this study, for the first time, we show how the electrical charge of small EVs can be more accurately determined by accounting for the impact of tightly bound ions. This was accomplished by measuring the electrophoretic mobility of EVs, and then analytically correlating the measured values to their charge in the form of zeta potential and electrical valence. In contrast to the currently used theoretical expressions, the employed analytical method in this study enabled a more accurate estimation of EV surface charge, which will facilitate the development of EV‐based diagnostic and therapeutic applications.

Funder

Natural Sciences and Engineering Research Council of Canada

Publisher

Wiley

Subject

Cell Biology,Histology

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