Nanoparticle Characterization by TRPS

The size distribution and particle concentration of fractions recovered after ODG were measured using tunable resistive pulse sensing (TRPS) by qNano (Izon, Christchurch, New Zealand) following the manufacturer’s instructions for working with smaller range nanopores. Voltage and pressure values were set to optimize the signal to ensure high sensitivity. A nanopore NP100 was used for all fractions analyzed except for the grape vesicles, where a NP150 was used. Calibration was performed using CP100 carboxylated polystyrene calibration particles (Izon) at a 1:1,000 dilution. Samples were diluted 1:5 and applied to the nanopore. The size and concentration of particles were determined using the software provided by Izon (version 3.2). Protein concentration was measured in all fractions, and EV purity determined as described previously (35 (link)).

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Eichenberger R.M., Ryan S., Jones L., Buitrago G., Polster R., Montes de Oca M., Zuvelek J., Giacomin P.R., Dent L.A., Engwerda C.R., Field M.A., Sotillo J, & Loukas A. (2018). Hookworm Secreted Extracellular Vesicles Interact With Host Cells and Prevent Inducible Colitis in Mice. Frontiers in Immunology, 9, 850.

Publication 2018

qNano CP100 carboxylated polystyrene calibration particles

Dilution Grape Polystyrene Pressure Protein Sensitivity

Corresponding Organization : Australian Institute of Tropical Health and Medicine

Other organizations : QIMR Berghofer Medical Research Institute, Queensland Health, University of Adelaide, Australian National University

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Variable analysis

independent variables

Voltage and pressure values set to optimize the signal

dependent variables

Size distribution of fractions
Particle concentration of fractions
Protein concentration in all fractions
EV purity

control variables

Nanopore NP100 used for all fractions except grape vesicles (NP150 used)
Calibration performed using CP100 carboxylated polystyrene calibration particles at a 1:1,000 dilution
Samples diluted 1:5 and applied to the nanopore

controls

Positive control: CP100 carboxylated polystyrene calibration particles
Negative control: Not explicitly mentioned

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