Investigate the efficacy of size exclusion chromatography for the isolation of extracellular vesicles from C. elegans.

Autor: Thomas S; Department of Zoology, Faculty of Sciences, University of Delhi, Delhi 110007, India., Kaur J; Department of Zoology, Faculty of Sciences, University of Delhi, Delhi 110007, India., Kamboj R; USIC, Faculty of Sciences, University of Delhi, Delhi 110007, India., Thangariyal S; Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences, Delhi 110070, India., Yadav R; Department of Chemistry, Indian Institute of Technology, New Delhi, Delhi 110016, India., Kumar K; Department of Chemistry, Kirori Mal College, University of Delhi, Delhi 110007, India., Dhania NK; Department of Zoology, Faculty of Sciences, University of Delhi, Delhi 110007, India. Electronic address: narender@zoology.du.ac.in.
Jazyk: angličtina
Zdroj: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2024 Feb 01; Vol. 1233, pp. 123982. Date of Electronic Publication: 2023 Dec 30.
DOI: 10.1016/j.jchromb.2023.123982
Abstrakt: Isolation of Extracellular Vesicles (EVs) has been done extensively in the past using ultracentrifugation, a recent shift has been observed towards precipitation, and exosome isolation kits. These methods often co-elute contaminants of similar size and density which makes their detection and downstream applications quite challenging. As well as the EV yield is also compromised in some methodologies due to aggregate formation. In recent reports, size-exclusion chromatography (SEC) is replacing density gradient-based ultracentrifugation as the gold standard of exosome isolation. It outperforms in yield, purity and does not account for any physical damage to the EVs. We have standardized the methodology for an efficient pure yield of homogenous exosomes of size even smaller than 75 nm in Caenorhabditis elegans homogenate. The paper entails the application and optimization of EV isolation by SEC based on previous studies by optimizing bed size and type of sepharose column employed. We propose that this method is economically feasible in comparison with currently available approaches. A comparative study was conducted to investigate the performance of CL-6B in relation to CL-2B and further, this was combined with ultracentrifugation for higher efficacy. The methodology could be introduced in a clinical setting due to its therapeutic potential and scope. The eluted EVs were studied by flow cytometry, nanotracking and characterized for size and morphology.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Dr Narender K Dhania reports financial support was provided by Science and Engineering Research Board. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.].
(Copyright © 2023 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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