Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

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Eskelin , K , Lampi , M , Meier , F , Moldenhauer , E , Bamford , D H & Oksanen , H M 2017 , ' Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation ' , Extremophiles , vol. 21 , no. 6 , pp. 1119-1132 . https://doi.org/10.1007/s00792-017-0963-x

Title: Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation
Author: Eskelin, Katri; Lampi, Mirka; Meier, Florian; Moldenhauer, Evelin; Bamford, Dennis H.; Oksanen, Hanna M.
Contributor: University of Helsinki, Biosciences
University of Helsinki, Biosciences
University of Helsinki, Biosciences
University of Helsinki, Biosciences
Date: 2017-11
Language: eng
Number of pages: 14
Belongs to series: Extremophiles
ISSN: 1431-0651
URI: http://hdl.handle.net/10138/298119
Abstract: Viruses come in various shapes and sizes, and a number of viruses originate from extremities, e.g. high salinity or elevated temperature. One challenge for studying extreme viruses is to find efficient purification conditions where viruses maintain their infectivity. Asymmetrical flow field-flow fractionation (AF4) is a gentle native chromatography-like technique for size-based separation. It does not have solid stationary phase and the mobile phase composition is readily adjustable according to the sample needs. Due to the high separation power of specimens up to 50 A mu m, AF4 is suitable for virus purification. Here, we applied AF4 for extremophilic viruses representing four morphotypes: lemon-shaped, tailed and tailless icosahedral, as well as pleomorphic enveloped. AF4 was applied to input samples of different purity: crude supernatants of infected cultures, polyethylene glycol-precipitated viruses and viruses purified by ultracentrifugation. All four virus morphotypes were successfully purified by AF4. AF4 purification of culture supernatants or polyethylene glycol-precipitated viruses yielded high recoveries, and the purities were comparable to those obtained by the multistep ultracentrifugation purification methods. In addition, we also demonstrate that AF4 is a rapid monitoring tool for virus production in slowly growing host cells living in extreme conditions.
Subject: Asymmetrical flow field-flow fractionation
Virus purification
Archaeal virus
Macromolecular complex
High ionic strength
Halophilic virus
ANGLE LIGHT-SCATTERING
ARCHAEAL VIRUSES
IONIC-STRENGTH
PARTICLE-SIZE
SEPARATION
FAMILY
HIS1
BACTERIOPHAGES
SEDIMENTATION
ENVIRONMENTS
1183 Plant biology, microbiology, virology
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