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MethodsX. 2020 Feb 20;7:100806. doi: 10.1016/j.mex.2020.100806. eCollection 2020.

Titration methods for rVSV-based vaccine manufacturing.

Author information

1
Department of Bioengineering, McGill University, 3480 University, Montréal, Québec H3A 0E9, Canada.
2
Human Health Therapeutics, National Research Council Canada, Montreal, QC, Canada.

Abstract

The recombinant Vesicular Stomatitis Virus (rVSV) is an emerging platform for viral vector-based vaccines. Promising results have been reported in clinical trials for the rVSV-ZEBOV vaccine for Ebola virus disease prevention. In this study, we describe the titration tools elaborated to assess the titre of rVSV-ZEBOV productions. • A streamlined Median Tissue Culture Infectious Dose (TCID50) assay to determine the infectious titer of this vaccine was established. • A digital polymerase chain reaction (dPCR) assay to assess the total number of viral particles present in cell-free culture supernatants of rVSV productions was developed. • These assays are used to titre rVSV-ZEBOV samples and characterize the ratio of total particles to infectious units for monitoring process robustness and product quality attributes and can be used to titre samples generated in the production of further rVSV vectors.

KEYWORDS:

Ebola; HIV; TCID50; dPCR; rVSV-HIV; rVSV-ZEBOV

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