Izvorni znanstveni članak
The Role of Isolation Methods on a Nanoscale Surface Structure and Its Effect on the Size of Exosomes
JungReem Woo
; Department of Chemistry and Biochemistry, University of California, Los Angeles, USA
Shivani Sharma
; California NanoSystems Institute, University of California, Los Angeles, California, USA
James Gimzewski
; Department of Chemistry and Biochemistry, University of California, Los Angeles, USA; California NanoSystems Institute, University of California, Los Angeles, California, USA
Sažetak
Exosomes are ~100 nanometre diameter vesicles secreted by mammalian cells. These emerging disease biomarkers carry nucleic acids, proteins and lipids specific to the parental cells that secrete them. Exosomes are typically isolated in bulk by ultracentrifugation, filtration or immunoaffinity precipitation for downstream proteomic, genomic, or lipidomic analysis. However, the structural properties and heterogeneity of isolated exosomes at the single vesicle level are not well characterized due to their small size. In this paper, by using high-resolution atomic force microscope imaging, we show the nanoscale morphology and structural heterogeneity in exosomes derived from U87 cells. Quantitative assessment of single exosomes reveals nanoscale variations in morphology, surface roughness and counts isolated by ultracentrifugation (UC) and immunoaffinity (IA) purification. Both methods produce intact globular, 30-120 nm sized vesicles when imaged under fluid and in air. However, IA exosomes had higher surface roughness and bimodal size population compared to UC exosomes. The study highlights the differences in size and surface topography of exosomes purified from a single cell type using different isolation methods.
Ključne riječi
AFM; Exosome; Nanoparticles; Immunoaffinity; Surface Roughness; Vesicles; Isolation Method
Hrčak ID:
161483
URI
Datum izdavanja:
1.1.2016.
Posjeta: 868 *