Affiliation:
1. School of Chemical Engineering The University of Queensland Brisbane QLD 4072 Australia
2. Translational Extracellular Vesicles in Obstetrics and Gynae‐Oncology Group Faculty of Medicine University of Queensland Centre for Clinical Research Royal Brisbane and Women's Hospital The University of Queensland Brisbane QLD 4029 Australia
3. Centre for Microscopy and Microanalysis The University of Queensland Brisbane QLD 4072 Australia
4. University of Melbourne Department of Obstetrics and Gynaecology Australia, and Mercy Hospital for Women 163 Studley Road Heidelberg Victoria 3084 Australia
5. Australian Institute of Bioengineering and Nanotechnology The University of Queensland Brisbane QLD 4072 Australia
6. Department of Nanomedicine Houston Methodist Research Institute Houston TX 77030 USA
Abstract
AbstractExtracellular vesicles (EVs) are nanosized biomolecular packages involved in intercellular communication. EVs are released by all cells, making them broadly applicable as therapeutic, diagnostic, and mechanistic components in (patho)physiology. Sample purity is critical for correctly attributing observed effects to EVs and for maximizing therapeutic and diagnostic performance. Lipoprotein contaminants represent a major challenge for sample purity. Lipoproteins are approximately six orders of magnitude more abundant in the blood circulation and overlap in size, shape, and density with EVs. This study represents the first example of an EV purification method based on the chemically‐induced breakdown of lipoproteins. Specifically, a styrene‐maleic acid (SMA) copolymer is used to selectively breakdown lipoproteins, enabling subsequent size‐based separation of the breakdown products from plasma EVs. The use of the polymer followed by tangential flow filtration or size‐exclusion chromatography results in improved EV yield, preservation of EV morphology, increased EV markers, and reduced contaminant markers. SMA‐based EV purification enables improved fluorescent labeling, reduces interactions with macrophages, and enhances accuracy, sensitivity, and specificity to detect EV biomarkers, indicating benefits for various downstream applications. In conclusion, SMA is a simple and effective method to improve the purity and yield of plasma‐derived EVs, which favorably impacts downstream applications.
Subject
Biomaterials,Biotechnology,General Materials Science,General Chemistry
Cited by
1 articles.
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