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Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation
- Source :
- Extremophiles : life under extreme conditions. 21(6)
- Publication Year :
- 2017
-
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.
- Subjects :
- 0301 basic medicine
Archaeal Viruses
Archaeal virus
viruses
Asymmetrical Flow Field-Flow Fractionation
BACTERIOPHAGES
Fractionation
Macromolecular complex
01 natural sciences
Microbiology
Virus
03 medical and health sciences
Virus purification
Microbial ecology
Halophilic virus
SEDIMENTATION
1183 Plant biology, microbiology, virology
Infectivity
Chromatography
Asymmetrical flow field-flow fractionation
Chemistry
010401 analytical chemistry
General Medicine
Salt Tolerance
PARTICLE-SIZE
Halophile
ANGLE LIGHT-SCATTERING
FAMILY
0104 chemical sciences
030104 developmental biology
IONIC-STRENGTH
SEPARATION
High ionic strength
Molecular Medicine
Ultracentrifuge
HIS1
ENVIRONMENTS
Subjects
Details
- ISSN :
- 14334909
- Volume :
- 21
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- Extremophiles : life under extreme conditions
- Accession number :
- edsair.doi.dedup.....586f8b21775a56e646894a5eaf1e275c