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

Carnell, George, Grehan, Keith, Ferrara, Francesca, Molesti, Eleonora, 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). Article Number 2514. ISSN 2331-8325. E-ISSN 2331-8325. (doi:10.21769/BioProtoc.2514) (KAR id:62472)

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
DOI/Identification number: 10.21769/BioProtoc.2514
Uncontrolled keywords: MERS coronavirus, Lentiviral pseudotype, Virus neutralisation
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Divisions > Division of Natural Sciences > Medway School of Pharmacy
Depositing User: Nigel Temperton
Date Deposited: 28 Jul 2017 14:07 UTC
Last Modified: 04 Mar 2024 16:33 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/62472 (The current URI for this page, for reference purposes)

University of Kent Author Information

Grehan, Keith.

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Ferrara, Francesca.

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Molesti, Eleonora.

Creator's ORCID:
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Temperton, Nigel J..

Creator's ORCID: https://orcid.org/0000-0002-7978-3815
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