Abstract
At present, there are no vaccines available for hand, foot, and mouth disease, which is caused by Coxsackie virus A16 (CVA16) infection. In the present study, we isolated epidemic strains of CVA16 and optimized the production of the virus in Vero cells. The system comprised growing the infected cells on polymer fiber paper carriers in a serum-free medium containing 0.5% (w/v) lactalbumin hydrolysate a mini bioreactor. Disposable Bioflo310 and AmProtein Current perfusion bioreactors were used to monitor virus infection and Vero cell culture. The total number of cells increased from 1.5 × 109 to 3.0 × 1010. In our optimized culture process, the virus titer reached 7.8 × 107 TCID50/mL at three days after infection. The inactivated CVA16 prepared from our optimized culture procedure elicited a slightly higher neutralizing antibody titer compared with that derived from routine culture procedures. These results will promote the large-scale production of inactivated CVA16 vaccines using nonwoven polymer fiber paper cell cultures.
Highlights
With the emergence of new infectious diseases in recent years, the technology of the large-scale culture of viruses has displayed important advantages, which is of great significance for the development of vaccines an antibodies
In Group 1 (G1), coxsackie virus A16 (CVA16) was inoculated into Vero cells in Dulbecco’s Modified Eagle Medium (DMEM) for 2 h, followed by supplementation using 10% fetal bovine serum (FBS)
In Group 2 (G2), CVA16 was inoculated into Vero cells in DMEM for 2 h, followed by the addition of virus production serum-free medium (VP-SFM)
Summary
With the emergence of new infectious diseases in recent years, the technology of the large-scale culture of viruses has displayed important advantages, which is of great significance for the development of vaccines an antibodies. In large-scale bioreactor systems, the cost of serum-free media is very high. To solve these problems, the present study developed a relatively serum-free culture system via a process that uses lactalbumin hydrolysate (LH) as a medium supplement during the harvest period. We used the AmProtein Current Perfusion Bioreactor (ACPB) system from AmProtein (Hangzhou, China) in 2009 [27] to create a cell/virus bank and optimized the culture conditions for the production of viruses. The optimized conditions were used in the ACPB and Bioflo310 bioreactors for CVA16 virus production [28]
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