Purification and proteomics of influenza virions

Hutchinson, E. C. and Stegmann, M. (2018) Purification and proteomics of influenza virions. In: Yamauchi, Y. (ed.) Influenza Virus. Series: Methods in molecular biology (1836). Humana Press: New York, NY, pp. 89-120. ISBN 9781493986774 (doi:10.1007/978-1-4939-8678-1_5)

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Abstract

This chapter describes a basic workflow for analyzing the protein composition of influenza virions. In order to obtain suitable material, the chapter describes how to concentrate influenza virions from the growth media of infected cells and to purify them by ultracentrifugation through a density gradient. This approach is also suitable for purifying influenza virions from the allantoic fluid of embryonated chicken eggs. As a small quantity of microvesicles are co-purified with virions, optional steps are included to increase the stringency of purification by enriching material with viral receptor binding and cleaving activity. Material purified in this way can be used for a variety of downstream applications, including proteomics. As a detailed example of this, the chapter also describes a standard workflow for analyzing the protein composition of concentrated virions by liquid chromatography and tandem mass spectrometry.

Item Type:Book Sections
Additional Information:Online ISBN: 9781493986781.
Status:Published
Glasgow Author(s) Enlighten ID:Hutchinson, Dr Edward
Authors: Hutchinson, E. C., and Stegmann, M.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Publisher:Humana Press
ISSN:1064-3745
ISBN:9781493986774
Published Online:28 August 2018
Copyright Holders:Copyright © 2018 Springer Science+Business Media B.V., part of Springer Nature
First Published:First published in Influenza Virus: 89-120
Publisher Policy:Reproduced in accordance with the publisher copyright policy

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
703851Functional investigations of the influenza virus proteomeEdward HutchinsonMedical Research Council (MRC)MR/N008618/1MVLS III - CENTRE FOR VIRUS RESEARCH