A new application of monolithic supports: the separation of viruses from one another.
The emergence of next-generation "deep" sequencing has enabled the study of virus populations with much higher resolutions. This new tool increases the possibility of observing mixed infections caused by combinations of plant viruses, which are likely to occur more frequently than previously thought. The biological impact of co-infecting viruses on their host has yet to be determined and fully understood, and the first step towards reaching this goal is the separation and purification of individual species. Ion-exchange monolith chromatography has been used successfully for the purification and concentration of different viruses, and number of them have been separated from plant homogenate or bacterial and eukaryotic lysate. Thus, the question remained as to whether different virus species present in a single sample could be separated. In this study, anion-exchange chromatography using monolithic supports was optimized for fast and efficient partial purification of three model plant viruses: Turnip yellow mosaic virus, Tomato bushy stunt virus, and Tobacco mosaic virus. The virus species, as well as two virus strains, were separated from each other in a single chromatographic experiment from an artificially mixed sample. Based on A260/280 ratios, we were able to attribute specific peaks to a certain viral morphology/structure (icosahedral or rod-shaped). This first separation of individual viruses from an artificially prepared laboratory mixture should encourage new applications of monolithic chromatographic supports in the separation of plant, bacterial, or animal viruses from all kinds of mixed samples.
Department of Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia. Electronic address: jelena.ruscic@biol.pmf.hr.
J
J
Ruščić
Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia.
I
I
Gutiérrez-Aguirre
Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia.
M Tušek
MT
Žnidarič
Department of Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
S
S
Kolundžija
Department of Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
A
A
Slana
BIA Separations d.o.o., Ajdovščina, Slovenia.
M
M
Barut
Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia.
M
M
Ravnikar
Department of Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
M
M
Krajačić
D002852:Chromatography, Ion Exchange / Q000379:methods*; D014027:Tobacco Mosaic Virus / Q000302:isolation & purification*; D017864:Tombusvirus / Q000302:isolation & purification*; D017806:Tymovirus / Q000302:isolation & purification*
Ion-exchange chromatography; Mixed infections; Monolithic supports; Plant viruses; Virus strains