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Biophysical analysis of BMV virions purified using a novel method

A. Strugała, M. Kręcisz, J. D. Rybka, A. Urbanowicz, K. Szpotkowski, P. Bierwagen, M. Figlerowicz, M. Kozak, C. Böttcher, M. Giersig – 2017

Brome mosaic virus (BMV) has been successfully loaded with different types of nanoparticles. However, studies concerning its application as a nanoparticle carrier demand high-purity virions in large amounts. Existing BMV purification protocols rely on multiple differential ultracentrifugation runs of the initially purified viral preparation. Herein, we describe an alternative method for BMV purification based on ion-exchange chromatography and size-exclusion chromatography (SEC) yielding 0.2mg of virus from 1g of plant tissue. Our method is of similar efficiency to previously described protocols and can easily be scaled up. The method results in high-quality BMV preparations as confirmed by biophysical analyses, including cryogenic transmission electron microscopy (cryo-TEM), dynamic light scattering (DLS), static light scattering (SLS), and circular dichroism (CD) measurements and attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) spectroscopy. Our results revealed that purified BMV capsids are stable and monodisperse and can be used for further downstream applications. In this work, we also characterize secondary structure and size fluctuations of the BMV virion at different pH values.

Titel
Biophysical analysis of BMV virions purified using a novel method
Verfasser
A. Strugała, M. Kręcisz, J. D. Rybka, A. Urbanowicz, K. Szpotkowski, P. Bierwagen, M. Figlerowicz, M. Kozak, C. Böttcher, M. Giersig
Datum
2017
Kennung
DOI: 10.1016/j.jchromb.2017.10.022
Zitierweise
J Chrom B 2017, 1068–1069, 157-163
Sprache
eng
Art
Text