Production of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using different culture techniques by vero cell line

SARS-CoV-2, which caused the World Health Organization to declare an international public health emergency, as of 30 November 2022, caused the death of more than 6.6 million people. While there are 50 approved vaccines worldwide against the disease, 11 of them have been approved by the WHO. According to the vaccine production data of 2021, 11.15 billion doses of vaccine were produced for Covid-19. Inactivated vaccines constitute approximately 45% of the total vaccine production. The first step in the inactivated vaccine process is the production of live virus. Optimization of SARS-CoV-2 production and purification is important to determine which system is efficient. The aim of this study was to investigate the production and purification of SARS-CoV-2 alpha variant antigen in high and low serum concentration by Vero cells in Cell Factory (CF) system for static culture and microcarrier (MC) spinner flask system for suspended culture. Antigen amount of pellets obtained by PEG precipitation of clarified culture was determined by SDS-PAGE. According to the calculations, higher efficiency was obtained at 5% FBS conditions during cell production and 1% during virus production. According to the first study results, antigen yields; It was obtained as 502.5 µg antigen/liter in the CF system and 355.9 µg antigen/liter in the MC system. For purification studies, the yield was increased to 756.7 µg antigen/liter in the second run with the CF system. Concentrated antigen was purified in AKTA pure-150 according to multimodal (Capto Core 400, CC400) and ion-exchange (Nuvia HP-Q) principles. As a result, yield values of 89% for CC400 and 74% for Nuvia HP-Q were found. Also, CC400 column gave better results in terms of removing impurities. As a result, the Cell Factory system was found to be more efficient for the laboratory-scale production of SARS-CoV-2 antigen and the CC400 column gave the best results for the purification process.