Development and scale-up of rVSV-SARS-CoV-2 vaccine process using single use bioreactor

Ton, Christopher; Stabile, Victoria; Carey, Elizabeth J.; Maraikar, Adam; Whitmer, Travis; Marrone, Samantha; Afanador, Nelson Lee; Zabrodin, Igor; Manomohan, Greeshma; Whiteman, Melissa C.; Hofmann, Carl

doi:10.1016/j.btre.2023.e00782

Cited by 12 publications

(12 citation statements)

References 38 publications

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“…Furthermore, incorporating intermediate equipment such as Harvestainer TM [ 50 ] or other mesh screens [ 25 ] to separate microcarriers from cells adds an additional transfer step at each scale, resulting in a more complex and costly process.…”

Section: Discussionmentioning

confidence: 99%

Seed Train Optimization in Microcarrier-Based Cell Culture Post In Situ Cell Detachment through Scale-Down Hybrid Modeling

Ebrahimian,

Schalk,

Dürkop

et al. 2024

Bioengineering

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Microcarrier-based cell culture is a commonly used method to facilitate the growth of anchorage-dependent cells like MA 104 for antigen manufacturing. However, conventionally, static cell culture is employed for cell propagation before seeding the production bioreactor with microcarriers (MCs). This study demonstrates the effective replacement of the conventional method by serial subculturing on MCs with in situ cell detachment under optimal conditions in closed culture units. This study proves that MA 104 can be subcultured at least five times on Cytodex 1 MC without the need for separating cells and MC after cell harvest. Process parameters impacting cell growth were studied post in situ cell detachment in a scaled-down model. Optimization, using augmented Design of Experiments (DoE) combined with hybrid modeling, facilitated rapid screening of the design space for critical process parameters (CPPs). Optimized conditions included an inoculation density of >16 cells/bead, 3.5–4.5 g/L of Cytodex 1, and a controlled agitation speed, starting at Njs (minimum agitation speed) for the first day with a maximum increase of 25% thereafter. With these design spaces for CPPs, a cell density of 2.6 ± 0.5 × 106 cells/mL was achieved after five days. This refined bioprocess methodology offers a reliable and efficient approach for seed training in stirred tank reactors, which is particularly beneficial for viral vaccine production.

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Section: Discussionmentioning

confidence: 99%

Seed Train Optimization in Microcarrier-Based Cell Culture Post In Situ Cell Detachment through Scale-Down Hybrid Modeling

Ebrahimian,

Schalk,

Dürkop

et al. 2024

Bioengineering

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“…As previously described by Ton et al., [ 20 ] Vero cells (ATCC CCL‐81.2) were expanded in Corning CellSTACKs (Corning, NY) for four passages and then planted into N‐1 single‐use bioreactors (ThermoFisher, MA).…”

Section: Methodsmentioning

confidence: 99%

“…As previously described by Ton et al, [20] Vero cells (ATCC CCL-81. C, dissolved oxygen was controlled to 50%, and pH was controlled at 7.3 for the duration of growth.…”

Section: Production Of Rvsvδg-sars-cov-2 Virusmentioning

confidence: 99%

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

Kuczynski,

Shallow,

Watson

et al. 2023

Biotechnology Journal

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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.

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“…Previous studies have reported the development of bioprocesses for VSV-based vaccines using Vero cells adhered to microcarriers, such as Cytodex 1 ( Kiesslich et al, 2020 ), or in suspension ( Kiesslich et al, 2021 ). More specifically, the production of the rVSV-ΔG-spike vaccine in Vero cells using Cytodex 1 has recently been reported ( Ton et al, 2023 ). However, these reports have indicated that further process improvement is needed to enhance titer yields.…”

Section: Introductionmentioning

confidence: 99%

Enhanced production yields of rVSV-SARS-CoV-2 vaccine using Fibra-Cel® macrocarriers

Cohen,

Simon,

Hazan

et al. 2024

Front. Bioeng. Biotechnol.

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The COVID-19 pandemic has led to high global demand for vaccines to safeguard public health. To that end, our institute has developed a recombinant viral vector vaccine utilizing a modified vesicular stomatitis virus (VSV) construct, wherein the G protein of VSV is replaced with the spike protein of SARS-CoV-2 (rVSV-ΔG-spike). Previous studies have demonstrated the production of a VSV-based vaccine in Vero cells adsorbed on Cytodex 1 microcarriers or in suspension. However, the titers were limited by both the carrier surface area and shear forces. Here, we describe the development of a bioprocess for rVSV-ΔG-spike production in serum-free Vero cells using porous Fibra-Cel® macrocarriers in fixed-bed BioBLU®320 5p bioreactors, leading to high-end titers. We identified core factors that significantly improved virus production, such as the kinetics of virus production, the use of macrospargers for oxygen supply, and medium replenishment. Implementing these parameters, among others, in a series of GMP production processes improved the titer yields by at least two orders of magnitude (2e9 PFU/mL) over previously reported values. The developed process was highly effective, repeatable, and robust, creating potent and genetically stable vaccine viruses and introducing new opportunities for application in other viral vaccine platforms.

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Development and scale-up of rVSV-SARS-CoV-2 vaccine process using single use bioreactor

Cited by 12 publications

References 38 publications

Seed Train Optimization in Microcarrier-Based Cell Culture Post In Situ Cell Detachment through Scale-Down Hybrid Modeling

Seed Train Optimization in Microcarrier-Based Cell Culture Post In Situ Cell Detachment through Scale-Down Hybrid Modeling

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

Enhanced production yields of rVSV-SARS-CoV-2 vaccine using Fibra-Cel® macrocarriers

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