Abstract
Due to their size, small extracellular vesicles such as exosomes have been difficult to identify and to quantify. As the roles that exosomes play in intercellular signalling become clearer, so does their potential utility as both diagnostic biomarkers for disease and as therapeutic vectors. Accurate assessment of exosomes, both their number and their cargo, is important for continued advancement in the field of vesicle research. To that end, several technologies, including nanoparticle tracking analysis, have been developed to define the physical characteristics of vesicle preparations and determine their concentration. This chapter describes a method for identifying the size and concentration of a subpopulation of vesicles in biological samples, using nanoparticle tracking analysis. Characterization of distinct exosomes is enabled by specific marker antibodies, coupled to fluorescent quantum dots.
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References
Skog J, Würdinger T, van Rijn S, Meijer DH, Gainche L, Sena-Esteves M, Curry WT Jr, Carter BS, Krichevsky AM, Breakefield XO (2008) Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nat Cell Biol 10:1470–1476
Colombo M, Raposo G, Théry C (2014) Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Ann Rev Cell Dev Biol 30:255–289
Gercel-Taylor C, Atay S, Tullis RH, Kesimer M, Taylor DD (2012) Nanoparticle analysis of circulating cell-derived vesicles in ovarian cancer patients. Anal Biochem 428:44–53
Sokolova V, Ludwig AK, Hornung S, Rotan O, Horn PA, Epple M, Giebel B (2011) Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. Colloids Surf B Biointerfaces 87:146–150
Dragovic RA, Gardiner C, Brooks AS, Tannetta DS, Ferguson DJ, Hole P, Carr B, Redman CW, Harris AL, Dobson PJ, Harrison P, Sargent IL (2011) Sizing and phenotyping of cellular vesicles using nanoparticle tracking analysis. Nanomedicine 7(6):780–788
Oosthuyzen W, Sime NE, Ivy JR, Turtle EJ, Street JM, Pound J, Bath LE, Webb DJ, Gregory CD, Bailey MA, Dear JW (2013) Quantification of human urinary exosomes by nanoparticle tracking analysis. J Physiol 591(23):5833–5842
Soo CY, Song Y, Zheng Y, Campbell EC, Riches AC, Gunn-Moore F, Powis SJ (2012) Nanoparticle tracking analysis monitors microvesicle and exosome secretion from immune cells. Immunology 136(2):192–197
King HW, Michael MZ, Gleadle JM (2012) Hypoxic enhancement of exosome release by breast cancer cells. BMC Cancer 12:421
Medintz IL, Uyeda HT, Goldman ER, Mattoussi H (2005) Quantum dot bioconjugates for imaging, labelling and sensing [review]. Nat Mater 4(6):435–446
Escola JM, Kleijmeer MJ, Stoorvogel W, Griffith JM, Yoshie O, Geuze HJ (1998) Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes. J Biol Chem 273:20121–20127
Heijnen HFG, Schiel AE, Fijnheer R, Geuze HJ, Sixma JJ (1999) Activated platelets release two types of membrane vesicles: microvesicles by surface shedding and exosomes derived from exocytosis of multivesicular bodies and alpha-granules. Blood 94:3791–3799
Thery C, Amigorena S, Raposo G, Clayton A (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol 3:22
Pisitkun T, Shen R-F, Knepper MA (2004) Identification and proteomic profiling of exosomes in human urine. Proc Natl Acad Sci U S A 101(36):13368–13373
Gardiner C, Ferreira YJ, Dragovic RA, Redman CW, Sargent IL (2013) Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis. J Extracell Vesicles 2:19671
Acknowledgments
Thanks to Rebecca Dragovic and NanoSight representatives for helpful discussions. Projects related to this chapter were funded by the Flinders Medical Centre Foundation and BioInnovation SA.
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McNicholas, K., Michael, M.Z. (2017). Immuno-characterization of Exosomes Using Nanoparticle Tracking Analysis. In: Hill, A. (eds) Exosomes and Microvesicles. Methods in Molecular Biology, vol 1545. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6728-5_3
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DOI: https://doi.org/10.1007/978-1-4939-6728-5_3
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