This technique is a novel method for preparing coxsackievirus vaccines by HEK293 cells. Serum-free HEK293 culture can be scaled up in attachedsuspension culture system,it can propagate various types of enteroviruses. The production of different enterovirus antigens can be included in a multivalent HFMD virus vaccine, which can be use to control the epidemic of multiple HFMD diseases.

Enteroviruses have different susceptibility to various cell lines due to their characteristics. Vero cell line is not a suitable host for the isolationpropagation of all enterovirus groups. Our method overcomes such limitsimproves the manufacturing of coxsackievirus vaccines using HEK293 cells. Serum-free HEK293 cells is a suitable host for various types of enteroviruses,reduce the risk of serum-containing medium for vaccine development.

This HEK293 cell preparation technique can be used to produce coxsackieviruses,can be utilized to develop multivalent HFMD vaccines. Coxsackievirus produced in serum-free HEK293 cells suspension culture system have higher virus production efficiency. The composition of different enterovirus antigens can be combined into a multivalent HFMD virus vaccine, which can be use to control the epidemic of multiple HFMD diseases.