RT Journal Article
T1 Isolation of goat milk small extracellular vesicles by novel combined bio-physical methodology
A1 González Vasco, María Isabel
A1 Gallardo Alguacil, Begoña
A1 Ceron, Carlos
A1 Aguilera-Jimenez, Elena
A1 Cortes-Canteli, Marta
A1 Peinado, Hector
A1 Desco Menéndez, Manuel
A1 Salinas Rodríguez, Beatriz
AB Introduction: Goat milk is notable as a cost-effective source of exosomes, also known as small extracellular vesicles (sEVs). These nanoparticle-like structures are naturally secreted by cells and have emerged as potential diagnostic agents and drug delivery systems, also supported by their proven therapeutic effects. However, the complexity of goat milk and the lack of standardized protocols make it difficult to isolate pure sEVs. This work presents an optimized approach that combines well-established physical isolation methods with the biological treatment of milk with rennet.Methods: sEVs derived from goat milk were purified using a methodology that combines differential ultracentrifugation, rennet, and size-exclusion chromatography. This novel strategy was compared with two of the main methodologies developed for isolating extracellular vesicles from bovine and human milk by means of physico-chemical characterization of collected vesicles using Transmission Electron Microscopy, Western blot, Bradford Coomassie assay, Dynamic Light Scattering, Nanoparticle Tracking Analysis and Zeta Potential.Results: Vesicles isolated with the optimized protocol had sEV-like characteristics and high homogeneity, while samples obtained with the previous methods were highly aggregated, with significant residual protein content.Discussion: This work provides a novel biophysical methodology for isolating highly enriched goat milk sEVs samples with high stability and homogeneity, for their further evaluation in biomedical applications as diagnostic tools or drug delivery systems.
PB Frontiers
SN 2296-4185
YR 2023
FD 2023-09-27
LK https://hdl.handle.net/10016/39710
UL https://hdl.handle.net/10016/39710
LA eng
NO This study has been funded by the Instituto de Salud Carlos III through the projects "PI20/01632" and "PT20/00044", and co-funded by European Regional Development Fund (ERDF, "A way to make Europe"), and the PRISMAP project. This work has been also funded by Comunidad de Madrid through the project "S2022/BMD-7403 RENIM-CM." The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia e Innovacion (MCIN), and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (CEX 2020-001041-S). MIG is funded by the Instituto de Investigacion Sanitaria Gregorio Maranon, Intramural Programme for the Promotion of R & D & I 2021, Sub-programme "Predoctoral training contract." MC-C is funded by a Miguel Servet type II research contract (CPII21/00007) from the Instituto de Salud Carlos III, Madrid, Spain. CC is funded by a predoctoral fellowship from Fundacion Espanola de Trombosis y Hemostasia (FETH-SETH).r This study has been funded by the Instituto de Salud Carlos III through the projects "PI20/01632" and "PT20/00044", and co-funded by European Regional Development Fund (ERDF, "A way to make Europe"), and the PRISMAP project. This work has been also funded by Comunidad de Madrid through the project "S2022/BMD-7403 RENIM-CM." The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia e Innovacion (MCIN), and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (CEX 2020-001041-S). MIG is funded by the Instituto de Investigacion Sanitaria Gregorio Maranon, Intramural Programme for the Promotion of R & D & I 2021, Sub-programme "Predoctoral training contract." MC-C is funded by a Miguel Servet type II research contract (CPII21/00007) from the Instituto de Salud Carlos III, Madrid, Spain. CC is funded by a predoctoral fellowship from Fundacion Espanola de Trombosis y Hemostasia (FETH-SETH).
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RD 30 jun. 2024