Adaptation of an rVSV Ebola vaccine purification process for rapid development of a viral vaccine candidate for SARS-CoV-2

Biotechnol J. 2024 Jan;19(1):e2300041. doi: 10.1002/biot.202300041. Epub 2023 Oct 10.


During the COVID-19 pandemic, long development timelines typically associated with vaccines were challenged. The urgent need for a vaccine provided a strong driver to reevaluate existing vaccine development approaches. Innovative approaches to regulatory approval were realized, including the use of platform-based technology. In collaboration with the International AIDS Vaccine Initiative, Inc. (IAVI), Merck & Co., Inc., Rahway, NJ, USA rapidly advanced an investigational SARS-CoV-2 vaccine based on the recombinant vesicular stomatitis virus (rVSV) platform used for the Ebola vaccine ERVEBO (rVSV∆G-ZEBOV-GP). An rVSV∆G-SARS-CoV-2 vaccine candidate was generated using the SARS-CoV-2 spike protein to replace the VSV G protein. The purification process development for this vaccine candidate was detailed in this paper. Areas were highlighted where the ERVEBO platform process was successfully adopted and where additional measures were needed for the SARS-CoV-2 vaccine candidate. These included: (i) endonuclease addition directly into the bioreactor prior to harvest, (ii) inclusion of a core-shell chromatography step for improved purification, and (iii) incorporation of a terminal, sterile filtration step to eliminate the need for aseptic, closed processing. High infectious virus titers were achieved in Phase 3 clinical drug substance (>108 PFU mL-1 ), and process consistency was demonstrated across four large scale batches that were completed in 6 months from clone selection.

Keywords: COVID-19; SARS-CoV-2; live virus purification; sterile filtration; vesicular stomatitis virus.

MeSH terms

  • Animals
  • Antibodies, Viral
  • COVID-19 Vaccines
  • COVID-19* / prevention & control
  • Ebola Vaccines* / genetics
  • Ebolavirus*
  • Hemorrhagic Fever, Ebola* / prevention & control
  • Humans
  • Pandemics
  • SARS-CoV-2 / genetics
  • Spike Glycoprotein, Coronavirus*
  • Vaccines, Synthetic
  • Vesicular Stomatitis*
  • Vesicular stomatitis Indiana virus
  • Vesiculovirus
  • Viral Vaccines*


  • Ebola Vaccines
  • Viral Vaccines
  • spike protein, SARS-CoV-2
  • COVID-19 Vaccines
  • Vaccines, Synthetic
  • Antibodies, Viral
  • Spike Glycoprotein, Coronavirus