Efficient strategy to isolate exosomes using anti-CD63 antibodies conjugated to gold nanoparticles

Abstract

Exosomes, a subpopulation of Extracellular vesicles (EVs), are cell-secreted vesicles found in the majority of biological fluids, including breast milk, tears, sweat, blood and, urine. The density and size of these vesicles depend on a variety of factors, including age, gender and the biological condition of the individual. Researchers are now focusing on the selective extraction of exosomes from bodily fluids due to the unique biomolecule composition of exosomes, which is critical for diagnosis, disease, and regeneration. Furthermore, current approaches for exosome isolation have limitations, necessitating the development of a simpler and more effective technique to achieve this goal. In this study, we investigated a quick and effective strategy for isolating exosomes from serum using a bench-top centrifuge. This was accomplished by raising antibodies against exosome surface tetraspanins (CD9, CD63 & CD81) in Leghorn chickens due to their phylogenetic distance from humans and cost-effectiveness for commercial use. In order to separate exosomes from a complex biological fluid, the antibodies were further coupled with gold nanoparticles (AuNPs). The findings were validated using ELISA, spectrophotometry, and transmission electron microscopy (TEM). Using this technique, exosome isolation from serum was achieved rapidly and these were captured by using anti CD63 antibodies bound to AuNPs. To summarize, exosomes were purified from serum using anti-CD63 antibodies conjugated to gold nanoparticles (IgY@AuNPs). Consequently, the approach for exosome isolation from biological fluid could be useful for clinically monitoring the biological state of the patients.

Recent research on material-based methods for isolation of extracellular vesicles

“…It was accomplished by employing antibodies targeting CD9, CD63 and CD81, and this approach for exosome isolation could be useful for clinically monitoring the biological state of the patients. 35…”

Classical and Innovative Evidence for Therapeutic Strategies in Retinal Dysfunctions

“…Exosomes might also be used in the field of gene therapy, as they can act as vehicles of therapeutic compounds like mRNA or siRNA. Attention is now focused on the use of exosomes as "therapeutic tools", as recent studies have analyzed the proteomic and transcriptomic profiles of these vesicles and demonstrated that they express molecules on their surface: some of these are found in all exosomes (LAMP2B and CD63) [98,99], while others are fabric-specific. This can be exploited specifically to deliver exosomes loaded with selective drugs to certain target cells [100].…”

“…proteomic and transcriptomic profiles of these vesicles and demonstrated that they express molecules on their surface: some of these are found in all exosomes (LAMP2B and CD63) [98,99], while others are fabric-specific. This can be exploited specifically to deliver exosomes loaded with selective drugs to certain target cells [100].…”