Production, purification, and capsid stability of rhinovirus C types

Journal of Virological Methods
Theodor F GriggsJames E Gern

Abstract

The rhinovirus C (RV-C) were discovered in 2006 and these agents are an important cause of respiratory morbidity. Little is known about their biology. RV-C15 (C15) can be produced by transfection of recombinant viral RNA into cells and subsequent purification over a 30% sucrose cushion, even though yields and infectivity of other RV-C genotypes with this protocol are low. The goal of this study was to determine whether poor RV-C yields were due to capsid instability, and moreover, to develop a robust protocol suitable for the purification of many RV-C types. Capsid stability assays indicated that virions of RV-C41 (refractory to purification) have similar tolerance for osmotic and temperature stress as RV-A16 (purified readily), although C41 is more sensitive to low pH. Modification to the purification protocol by removing detergent increased the yield of RV-C. Addition of nonfat dry milk to the sucrose cushion increased the virus yield but sacrificed purity of the viral suspension. Analysis of virus distribution following centrifugation indicated that the majority of detectable viral RNA (vRNA) was found in pellets refractory to resuspension. Reduction of the centrifugal force with commiserate increase in spin-time improved th...Continue Reading

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Citations

Aug 12, 2016·Proceedings of the National Academy of Sciences of the United States of America·Yue LiuMichael G Rossmann
Feb 26, 2019·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yong ZhangMichael J Holtzman
Sep 19, 2019·American Journal of Respiratory Cell and Molecular Biology·Vishal ParikhCynthia J Koziol-White
Jan 20, 2018·Emerging Infectious Diseases·Erik J ScullyTony L Goldberg
May 6, 2017·Respiratory Research·Theodor F GriggsJames E Gern
Dec 29, 2020·American Journal of Respiratory and Critical Care Medicine·Timothy ChoiYury A Bochkov
Nov 3, 2021·Archives of Virology·Marcilio Jorge FumagalliLuiz Tadeu Moraes Figueiredo

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