Residual matrix from different separation techniques impacts exosome biological activity View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-09

AUTHORS

Lucia Paolini, Andrea Zendrini, Giuseppe Di Noto, Sara Busatto, Elisabetta Lottini, Annalisa Radeghieri, Alessandra Dossi, Andrea Caneschi, Doris Ricotta, Paolo Bergese

ABSTRACT

Exosomes are gaining a prominent role in research due to their intriguing biology and several therapeutic opportunities. However, their accurate purification from body fluids and detailed physicochemical characterization remain open issues. We isolated exosomes from serum of patients with Multiple Myeloma by four of the most popular purification methods and assessed the presence of residual contaminants in the preparations through an ad hoc combination of biochemical and biophysical techniques - including Western Blot, colloidal nanoplasmonics, atomic force microscopy (AFM) and scanning helium ion microscopy (HIM). The preparations obtained by iodixanol and sucrose gradients were highly pure. To the contrary, those achieved with limited processing (serial centrifugation or one step precipitation kit) resulted contaminated by a residual matrix, embedding the exosomes. The contaminated preparations showed lower ability to induce NfkB nuclear translocation in endothelial cells with respect to the pure ones, probably because the matrix prevents the interaction and fusion of the exosomes with the cell membrane. These findings suggest that exosome preparation purity must be carefully assessed since it may interfere with exosome biological activity. Contaminants can be reliably probed only by an integrated characterization approach aimed at both the molecular and the colloidal length scales. More... »

PAGES

23550

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/srep23550

DOI

http://dx.doi.org/10.1038/srep23550

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1025955500

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/27009329


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