Development of an Integrated Continuous Manufacturing Process for the rVSV-Vectored SARS-CoV-2 Candidate Vaccine

• Published in: Vaccines (Basel) 2023-04, Vol.11 (4), p.841
• Main Authors: Yang, Zeyu, Paes, Barbara Cristina Martins Fernandes, Fulber, Julia Puppin Chaves, Tran, Michelle Yen, Farnós, Omar, Kamen, Amine A
• Format: Article
• Language: English
• Subjects: Air flow; Anion exchange; Anion exchanging; Bioreactors; Cell culture; Chromatography; Coronaviruses; COVID-19; COVID-19 vaccines; Deoxyribonuclease; Design of experiments; Efficiency; Genetic vectors; Immunization; Infections; Manufacturing; Manufacturing industry; Membranes; Modular systems; Pandemics; perfusion; Process parameters; Production processes; Proteins; recombinant vesicular stomatitis virus; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; Spike protein; Stomatitis; Temperature; Upstream; upstream process; Vaccines; Viral diseases; viral vectored vaccine
• ISSN: 2076-393X; 2076-393X
• DOI: 10.3390/vaccines11040841

The administration of viral vectored vaccines remains one of the most effective ways to respond to the ongoing novel coronavirus disease 2019 (COVID-19) pandemic. However, pre-existing immunity to the viral vector hinders its potency, resulting in a limited choice of viral vectors. Moreover, the basic batch mode of manufacturing vectored vaccines does not allow one to cost-effectively meet the global demand for billions of doses per year. To date, the exposure of humans to VSV infection has been limited. Therefore, a recombinant vesicular stomatitis virus (rVSV), which expresses the spike protein of SARS-CoV-2, was selected as the vector. To determine the operating upstream process conditions for the most effective production of an rVSV-SARS-CoV-2 candidate vaccine, a set of critical process parameters was evaluated in an Ambr 250 modular system, whereas in the downstream process, a streamlined process that included DNase treatment, clarification, and a membrane-based anion exchange chromatography was developed. The design of the experiment was performed with the aim to obtain the optimal conditions for the chromatography step. Additionally, a continuous mode manufacturing process integrating upstream and downstream steps was evaluated. rVSV-SARS-CoV-2 was continuously harvested from the perfusion bioreactor and purified by membrane chromatography in three columns that were operated sequentially under a counter-current mode. Compared with the batch mode, the continuous mode of operation had a 2.55-fold increase in space-time yield and a reduction in the processing time by half. The integrated continuous manufacturing process provides a reference for the efficient production of other viral vectored vaccines.