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J Virol Methods. 2012 Oct;185(1):166-70. doi: 10.1016/j.jviromet.2012.06.014. Epub 2012 Jun 26.

A plate-based high-throughput assay for virus stability and vaccine formulation.

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1
Division of Structural Biology, University of Oxford, The Henry Wellcome Building for Genomic Medicine, Roosevelt Drive, Oxford, OX3 7BN, UK.

Abstract

Standard methods for assessing the thermal stability of viruses can be time consuming and rather qualitative yet such data is a necessary requisite for vaccine formulation. In this study a novel plate-based thermal scanning assay for virus particle stability has been developed (PaSTRy: Particle Stability Thermal Release Assay). Two environment-sensitive fluorescent dyes, with non-overlapping emission spectra and different affinities, are used to accrue simultaneously independent data for the overall stability of the virus capsid, as judged by the exposure of the genome, and for capsid protein stability according to the exposure of hydrophobic side chains which are normally buried. This offers a fast and efficient high-throughput method to optimise vaccine formulation and to investigate the processes of virus uncoating.

PMID:
22744000
PMCID:
PMC3470038
DOI:
10.1016/j.jviromet.2012.06.014
[Indexed for MEDLINE]
Free PMC Article
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