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Table 3 Comparisons of isolation methods

From: Microparticles: biogenesis, characteristics and intervention therapy for cancers in preclinical and clinical research

Method Mechanism Pros Cons
Centrifugation Stepwise centrifugation: removing cells and debris at a Low centrifugation speed (300–1000 g), followed by MPs collection at a higher one (10000–20000 g) The most common and efficient isolation Method
Low cost
Simple to operate
Not easily contaminated
Wide range of sample volumes from a few millilitres up to > 100 mL
Low selectivity
Risk of aggregation and deformation of MPs
Risk of cosedimentation of larger vesicles and protein aggregates
Size exclusion chromatography By using a column packed with porous gels, large EVs flow out first than small EVs. Each component is separated according to size Reducing aggregation of MPs and proteins
Maintaining integrity and biological activity of MPs
Relatively high purification and inexpensive
In most cases, samples are diluted and re-concentration are required, resulting in long times
If there are multiple production cycles, columns need to be washed, sanitized and rebalanced
Ultrafiltration Use membranes with specific aperture to remove other components from the sample and retain and concentrate the MPs Low cost
Simple to operate
Easy for large scale
Low selectivity
Risks of non-specific binding of MPs to membranes and leading to some loss of yield
Risks of deformation or rupture of MPs
Immunoaffinity chromatography MPs are separated by the specific interaction of antigens on the surface of MPs and antibodies On the beads High purification
Capacity of isolating and quantifying specific sub-population of MPs
High cost
Unsuitable for large-volume samples and uneasy for large scale
Risks of deformation while elution
Accurately sorting a particular type of MPs, thus losing the heterogeneity of MPs
Microfluidics The isolation can be based on several aspects, such as shape, size, density, electric charge, specific lipid/proteins on MPs membrane High purification
Fast and simple to operate
Capacity of isolating and quantifying specific sub-population of MPs in real time
Available for small volume samples
Accurately sorting a particular type of MPs, thus losing the heterogeneity of MPs
Risks of deformation on account of the shear stress