Titration methods for rVSV-based vaccine manufacturing

Jean-François Gélinas,Sascha Kiesslich,Rénald Gilbert,Amine A Kamen

doi:10.1016/j.mex.2020.100806

Jean-François Gélinas, Sascha Kiesslich + Show 2 more

Open Access

https://doi.org/10.1016/j.mex.2020.100806

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Journal: MethodsX	Publication Date: Jan 1, 2020
Citations: 14	License type: cc-by

Affiliation: McGill University

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.

Highlights

The recombinant Vesicular Stomatitis Virus platform is a replication-competent vaccine that has been shown to generate both cell-mediated and humoral immunity to expressed foreign antigens [1] and is being developed to target several infectious diseases and cancers [2]
We describe the titration tools elaborated to assess the titre of recombinant Vesicular Stomatitis Virus (rVSV)-ZEBOV productions
This vaccine is based on the rVSV-ZEBOV, an attenuated, replication-competent rVSV pseudotyped with the Ebola Zaire glycoprotein [5]

Summary

Introduction

The recombinant Vesicular Stomatitis Virus (rVSV) platform is a replication-competent vaccine that has been shown to generate both cell-mediated and humoral immunity to expressed foreign antigens [1] and is being developed to target several infectious diseases and cancers [2]. The Ebola virus vaccine Ervebo has been approved by the FDA [3], prequalified by the World Health Organization and granted a conditional marketing by the European Commission [4]. This vaccine is based on the rVSV-ZEBOV, an attenuated, replication-competent rVSV pseudotyped with the Ebola Zaire glycoprotein [5]. The combination of a functional assay with a total particle assay through their ratio (functional particles over total particles) is a critical quality attribute for the characterization of the candidate rVSV-based vaccine as defective interfering rVSV particles have been shown to modulate virulence [6]

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Results

Conclusion