Evaluation of a downstream process for the recovery and concentration of a Cell-Culture-Derived rVSV-Spike COVID-19 vaccine candidate

Makovitzki, Arik; Lerer, Elad; Kafri, Yaron; Adar, Yaakov; Cherry, Lilach; Lupu, Edith; Monash, Arik; Levy, Rona; Israeli, Ofir; Dor, Eyal; Epstein, Eyal; Levin, Lilach; Toister, Einat; Hefetz, Idan; Hazan, Ophir; Simon, I. Hay; Tal, Arnon; Girshengorn, Meni; Tzadok, Hanan; Rosen, Osnat; Ziv, Oren

doi:10.1016/j.vaccine.2021.10.045

Cited by 9 publications

(11 citation statements)

References 31 publications

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“…The harvest solution was treated with nuclease, and a reduction of 99.9% in DNA concentration was achieved [ 26 ]. Host cell debris, large aggregates, and insoluble contaminants were clarified using a filter train composed of 3 µm and 1.2 µm inert polypropylene depth filters and a 0.45/0.2 µm membrane filter.…”

Section: Resultsmentioning

confidence: 99%

“…Throughout the clarification process, approximately 85% viral recovery was attained. Ultrafiltration and 4–5-fold diafiltration were performed with a 750 kDa HF against equilibration buffer, resulting in virus recovery of 45–55% [ 26 ]. The composition of the buffers used in this report, namely Tris and D-trehalose, is based on their importance as stabilizers in biological therapeutic products [ 58 , 59 ], specifically for the production of rVSV [ 30 , 60 ].…”

Section: Resultsmentioning

confidence: 99%

“…The DSP of cell culture-derived viral vaccines involves two discrete operations. The initial purification entails endonuclease digestion of the harvested supernatant, clarification, and concentration of the residual viral vector [ 26 ]. The final purification step uses chromatography to meet purity requirements [ 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

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Highly Efficient Purification of Recombinant VSV-∆G-Spike Vaccine against SARS-CoV-2 by Flow-Through Chromatography

Lerer

Ziv

Kafri

et al. 2021

BioTech

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This study reports a highly efficient, rapid one-step purification process for the production of the recombinant vesicular stomatitis virus-based vaccine, rVSV-∆G-spike (rVSV-S), recently developed by the Israel Institute for Biological Research (IIBR) for the prevention of COVID-19. Several purification strategies are evaluated using a variety of chromatography methods, including membrane adsorbers and packed-bed ion-exchange chromatography. Cell harvest is initially treated with endonuclease, clarified, and further concentrated by ultrafiltration before chromatography purification. The use of anion-exchange chromatography in all forms results in strong binding of the virus to the media, necessitating a high salt concentration for elution. The large virus and spike protein binds very strongly to the high surface area of the membrane adsorbents, resulting in poor virus recovery (<15%), while the use of packed-bed chromatography, where the surface area is smaller, achieves better recovery (up to 33%). Finally, a highly efficient chromatography purification process with CaptoTM Core 700 resin, which does not require binding and the elution of the virus, is described. rVSV-S cannot enter the inner pores of the resin and is collected in the flow-through eluent. Purification of the rVSV-S virus with CaptoTM Core 700 resulted in viral infectivity above 85% for this step, with the efficient removal of host cell proteins, consistent with regulatory requirements. Similar results were obtained without an initial ultrafiltration step.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Highly Efficient Purification of Recombinant VSV-∆G-Spike Vaccine against SARS-CoV-2 by Flow-Through Chromatography

Lerer

Ziv

Kafri

et al. 2021

BioTech

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“…The VSV-ΔG-spike vaccine was produced in BioBLU® 5p Eppendorf single-use bioreactors in Vero cell that were absorbed to Fibra-Cels. The medium from the bioreactors, containing the released vaccine virus, was harvested and transferred to downstream process (DSP) that comprises several steps (as described earlier ( Lerer et al, 2021 ; Makovitzki et al, 2021 ). The purification steps include: endonuclease digestion of host cell DNA, followed by clarification by a train of decreasing cut off filters, chromatographic purification, and tangential flow filtration (TFF).…”

Section: Methodsmentioning

confidence: 99%

SARS-CoV-2 spike antigen quantification by targeted mass spectrometry of a virus-based vaccine

Rosen

Jayson

Dor

et al. 2022

Journal of Virological Methods

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“…The DSP must eliminate impurities, either process-related (e.g., DNase, nutrients, extractables and leachables) or product-related (e.g., host cell proteins (HCP), metabolites and host cell DNA (hc-DNA)). The DSP that was developed by the IIBR for the rVSV-∆G-spike vaccine is a multistep process that comprised endonuclease digestion, depth and membrane filter clarification, chromatographic purification and ultrafiltration [ 9 , 10 ].…”

Section: Introductionmentioning

confidence: 99%

In-Line Monitoring of Downstream Purification Processes for VSV Based SARS-CoV-2 Vaccine Using a Novel Technique

Makovitzki

Jayson

Ziv

et al. 2021

BioTech

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The COVID-19 pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) increases the need for a rapid development of efficient vaccines. Among other vaccines in clinical trials, a recombinant VSV-∆G-spike vaccine was developed by the Israel Institute for Biological Research (IIBR) and is being evaluated. The development of an efficient downstream purification process (DSP) enables the vaccine to be advanced to clinical trials. The DSP must eliminate impurities, either process- or product-related, to yield a sufficient product with high purity, potency and quality. To acquire critical information on process restrictions and qualities, the application of in-line monitoring is vital and should significantly impact the process yield, product quality and economy of the entire process. Here, we describe an in-line monitoring technique that was applied in the DSP of the VSV-∆G-spike vaccine. The technique is based on determining the concentrations of metabolites, nutrients and a host cell protein using the automatic chemistry analyzer, Cobas Integra 400 Plus. The analysis revealed critical information on process parameters and significantly impacted purification processes. The technique is rapid, easy and efficient. Adopting this technique during the purification process improves the process yield and the product quality and enhances the economy of the entire downstream process for biotechnology and bio pharmaceutical products.

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Evaluation of a downstream process for the recovery and concentration of a Cell-Culture-Derived rVSV-Spike COVID-19 vaccine candidate

Cited by 9 publications

References 31 publications

Highly Efficient Purification of Recombinant VSV-∆G-Spike Vaccine against SARS-CoV-2 by Flow-Through Chromatography

Highly Efficient Purification of Recombinant VSV-∆G-Spike Vaccine against SARS-CoV-2 by Flow-Through Chromatography

SARS-CoV-2 spike antigen quantification by targeted mass spectrometry of a virus-based vaccine

In-Line Monitoring of Downstream Purification Processes for VSV Based SARS-CoV-2 Vaccine Using a Novel Technique

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