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2014 ; 406
(30
): 7855-66
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A review of exosome separation techniques and characterization of B16-F10 mouse
melanoma exosomes with AF4-UV-MALS-DLS-TEM
#MMPMID25084738
Petersen KE
; Manangon E
; Hood JL
; Wickline SA
; Fernandez DP
; Johnson WP
; Gale BK
Anal Bioanal Chem
2014[Dec]; 406
(30
): 7855-66
PMID25084738
show ga
Exosomes participate in cancer metastasis, but studying them presents unique
challenges as a result of their small size and purification difficulties.
Asymmetrical field flow fractionation with in-line ultraviolet absorbance,
dynamic light scattering, and multi-angle light scattering was applied to the
size separation and characterization of non-labeled B16-F10 exosomes from an
aggressive mouse melanoma cell culture line. Fractions were collected and further
analyzed using batch mode dynamic light scattering, transmission electron
microscopy and compared with known size standards. Fractogram peak positions and
computed radii show good agreement between samples and across fractions.
Ultraviolet absorbance fractograms in combination with transmission electron
micrographs were able to resolve subtle heterogeneity of vesicle retention times
between separate batches of B16-F10 exosomes collected several weeks apart.
Further, asymmetrical field flow fractionation also effectively separated B16-F10
exosomes into vesicle subpopulations by size. Overall, the flow field flow
fractionation instrument combined with multiple detectors was able to rapidly
characterize and separate exosomes to a degree not previously demonstrated. These
approaches have the potential to facilitate a greater understanding of exosome
function by subtype, as well as ultimately allow for "label-free" isolation of
large scale clinical exosomes for the purpose of developing future exosome-based
diagnostics and therapeutics.
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