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Gerritzen MJH, Martens DE, Wijffels RH, et al. High throughput nanoparticle tracking analysis for monitoring outer membrane vesicle production. J Extracell Vesicles. 2017;6(1):1333883doi: 10.1080/20013078.2017.1333883.
Gerritzen, M. J. H., Martens, D. E., Wijffels, R. H., & Stork, M. (2017). High throughput nanoparticle tracking analysis for monitoring outer membrane vesicle production. Journal of extracellular vesicles, 6(1), 1333883. https://doi.org/10.1080/20013078.2017.1333883
Gerritzen, Matthias J H, et al. "High throughput nanoparticle tracking analysis for monitoring outer membrane vesicle production." Journal of extracellular vesicles vol. 6,1 (2017): 1333883. doi: https://doi.org/10.1080/20013078.2017.1333883
Gerritzen MJH, Martens DE, Wijffels RH, Stork M. High throughput nanoparticle tracking analysis for monitoring outer membrane vesicle production. J Extracell Vesicles. 2017 Jun 19;6(1):1333883. doi: 10.1080/20013078.2017.1333883. eCollection 2017. PMID: 28717425; PMCID: PMC5505008.
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J Extracell Vesicles. 2017 Jun 19;6(1):1333883. doi: 10.1080/20013078.2017.1333883. eCollection 2017.

High throughput nanoparticle tracking analysis for monitoring outer membrane vesicle production.

Journal of extracellular vesicles

Matthias J H Gerritzen, Dirk E Martens, René H Wijffels, Michiel Stork

Affiliations

  1. Process Development Bacterial Vaccines, Institute for Translational Vaccinology (Intravacc), Bilthoven, The Netherlands.
  2. Bioprocess Engineering, Wageningen University, Wageningen, The Netherlands.
  3. Faculty of Biosciences and Aquaculture, Nord University, Bodø, Norway.

PMID: 28717425 PMCID: PMC5505008 DOI: 10.1080/20013078.2017.1333883

Abstract

Outer membrane vesicles (OMVs) are spherical membrane nanoparticles released by Gram-negative bacteria. OMVs can be quantified in complex matrices by nanoparticle tracking analysis (NTA). NTA can be performed in static mode or with continuous sample flow that results in analysis of more particles in a smaller time-frame. Flow measurements must be performed manually despite the availability of a sample changer on the NanoSight system. Here we present a method for automated measurements in flow mode. OMV quantification in flow mode results in lower variance in particle quantification (coefficient of variation (CV) of 6%, CV static measurements of 14%). Sizing of OMVs was expected to be less favorable in flow mode due to the increased movement of the particles. However, we observed a CV of 3% in flow mode and a CV of 8% in static measurements. Flow rates of up to 5 µL/min displayed correct size and particle measurements, however, particle concentration was slightly lower than in static measurements. The automated method was used to assess OMV release of batch cultures of

Keywords: NanoSight system; Nanoparticle tracking analysis; Neisseria meningitidis; autosampler; outer membrane vesicles; sample changer; syringe pump; vaccine production

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