Rebecca E Lane, Will Anderson, Deborah Hayoun, Darren Korbie, Matt Trau*

 

Australian Institute for Bioengineering and Nanotechnology
University of Queensland
St Lucia, QLD, Australia

 

Extracellular vesicles (EVs) including exosomes and microvesicles have garnered much recent interest for their potential utility as cancer biomarkers and therapeutic targets. EVs are membranous vesicles shed from cells into circulating bodily fluids, and are thought to act as a cell-cell communication mechanism. Although there is a substantial body of literature concerned with the protein and nucleic acid cargo of these vesicles, their physical properties remain relatively poorly understood. EVs exist primarily in the sub-200nm range and are physically heterogenous, and they therefore represent a complex and challenging sample for physical characterisation.

This presentation will discuss the use of four nanoparticle characterisation platforms: dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), tunable resistive pulse sensing (TRPS), and laser diffraction. Advantages and limitations of the various platforms with respect to EV characterisation will be discussed, including resolution of heterogeneity, ease of use and measurement time, limits of detection and measurement-to-measurement reproducibility. A particular focus will be our in-depth optimisation of the TRPS platform, and the measurement guidelines we have developed to improve system sensitivity and stability when measuring EV samples 2.   

The presentation will then highlight two applications of these platforms in EV research. The first is a comparative study of several methods for EV purification from biological fluids3. The second is the evaluation of synthetic liposomes as a model system for optimizing EV workflows. In total, it is anticipated that this work will enable a greater awareness and understanding of the particular challenges associated with the physical characterization of EVs, leading to improved experimental design in future investigations.  

References

1Colombo, M, Raposo, G, Théry, C. Annu. Rev. Cell Dev. Biol,. 2014, 30, 255-289. Biogenesis, Secretion, and Intercellular Interactions of Exosomes and Other Extracellular Vesicles.

2  Anderson, W, Lane, RE, Korbie, D, Trau M. Langmuir,  2015, 31, 6577-6587. Observations of Tunable Resistive Pulse Sensing for Exosome Analysis: Improving System Sensitivity and Stability.

3  Lane, RE, Korbie, D, Anderson, W, Vaidyanathan, R, Trau, M.  Sci Rep, 2015, 5, 7639. Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing.

 

Biographic Details

Name: Rebecca Lane             

Title: PhD Candidate

Affiliation, Country: Australian Institute for Bioengineering and Nanotechnology, Australia E-mail: r.lane2@uq.edu.au

Research interests: extracellular vesicles, cancer biomarkers