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

Autor: George Carnell, Keith Grehan, Francesca Ferrara, Eleonora Molesti, Nigel Temperton
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Bio-Protocol, Vol 7, Iss 16 (2017)
Druh dokumentu: article
ISSN: 2331-8325
DOI: 10.21769/BioProtoc.2514
Popis: 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.
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