An Optimised Method for the Production using PEI, Titration and Neutralization of SARS-CoV Spike Luciferase Pseudotypes

Carnell, George and Grehan, Keith and Ferrara, Francesca and Molesti, Eleonora and Temperton, Nigel J. (2017) An Optimised Method for the Production using PEI, Titration and Neutralization of SARS-CoV Spike Luciferase Pseudotypes. Bio-Protocol, 7 (16). ISSN 2331-8325. E-ISSN 2331-8325. (doi:https://doi.org/10.21769/BioProtoc.2514) (Full text available)

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Abstract

The protocol outlined represents a cost-effective, rapid and reliable method for the generation of high-titre viral pseudotype particles with the wild-type SARS-CoV spike protein on a lentiviral vector core using the widely available transfection reagent PEI. This protocol is optimized for transfection in 6-well plates; however it can be readily scaled to different production volumes according to application. This protocol has multiple benefits including the use of readily available reagents, consistent, high pseudotype virus production Relative Luminescence Units (RLU) titres and rapid generation of novel coronavirus pseudotypes for research into strain variation, tropism and immunogenicity/sero-prevalence.

Item Type: Article
Uncontrolled keywords: MERS coronavirus, Lentiviral pseudotype, Virus neutralisation
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Nigel Temperton
Date Deposited: 28 Jul 2017 14:07 UTC
Last Modified: 23 Aug 2017 13:14 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/62472 (The current URI for this page, for reference purposes)
Temperton, Nigel J.: https://orcid.org/0000-0002-7978-3815
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