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Updated: Dec 25, 2025

Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses
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Titration methods for rVSV-based vaccine manufacturing.

Jean-François Gélinas1, Sascha Kiesslich1, Rénald Gilbert1,2

  • 1Department of Bioengineering, McGill University, 3480 University, Montréal, Québec H3A 0E9, Canada.

Methodsx
|March 21, 2020
PubMed
Summary

New assays were developed to measure the infectious titer and total viral particles for the recombinant Vesicular Stomatitis Virus (rVSV) Ebola vaccine. These tools ensure robust production and quality control for viral vector vaccines.

Keywords:
EbolaHIVTCID50dPCRrVSV-HIVrVSV-ZEBOV

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Area of Science:

  • Virology
  • Vaccinology
  • Biotechnology

Background:

  • The recombinant Vesicular Stomatitis Virus (rVSV) is a promising viral vector platform for vaccine development.
  • The rVSV-ZEBOV vaccine has shown promising results in clinical trials for preventing Ebola virus disease.

Purpose of the Study:

  • To establish and describe titration tools for assessing the quantity of rVSV-ZEBOV productions.
  • To ensure robust production and quality control for viral vector vaccines.

Main Methods:

  • A streamlined Median Tissue Culture Infectious Dose (TCID50) assay was established to determine the infectious titer.
  • A digital polymerase chain reaction (dPCR) assay was developed to quantify total viral particles in cell-free supernatants.
  • These assays were used to titrate rVSV-ZEBOV samples and characterize the ratio of total particles to infectious units.

Main Results:

  • Established a streamlined TCID50 assay for infectious titer determination.
  • Developed a dPCR assay for quantifying total viral particles.
  • Characterized the ratio of total particles to infectious units for rVSV-ZEBOV samples.

Conclusions:

  • The developed titration assays are essential for monitoring process robustness and product quality attributes.
  • These methods can be applied to titrate samples from the production of other rVSV vectors.
  • This work supports the advancement of rVSV-based vaccine production and quality control.