Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 534177
Title High throughput nanoparticle tracking analysis for monitoring outer membrane vesicle production
Author(s) Gerritzen, Matthias J.H.; Martens, Dirk E.; Wijffels, René H.; Stork, Michiel
Source Journal of Extracellular Vesicles 6 (2017)1. - ISSN 2001-3078
DOI http://dx.doi.org/10.1080/20013078.2017.1333883
Department(s) LR - Animal Behaviour & Welfare
WIAS
Bioprocess Engineering
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2017
Keyword(s) autosampler - Nanoparticle tracking analysis - NanoSight system - Neisseria meningitidis - outer membrane vesicles - sample changer - syringe pump - vaccine production
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 Neisseria meningitidis. The bacteria released more OMVs in stationary growth phase, while the size of the vesicles remained constant throughout the culture. Taken together, this study shows that automated measurements in flow mode can be established with advanced scripting to reduce the workload for the user.
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