AbstractEnterovirus 71 (EV71) is an important agent responsible for hand‐foot‐and‐mouth disease (HFMD), which can cause severe neurological complications and death in children. However, there is no specific treatment for EV71 infection, and a safe and effective vaccine is needed urgently. In this study, an effective and economical method for the production of EV71‐VP1 protein was developed, and the VP1 protein was evaluated in humoral and cellular immune responses as an EV71 vaccine. The results revealed that the VP1 protein induced high titers of cross‐neutralizing antibodies for different EV71 subtypes, and elicited significant splenocyte proliferation. The high levels of IFN‐r and IL‐10 showed the VP1 protein induced a mixed Th1 and Th2 immune response. Vaccinated female mice could confer protection in their neonatal offspring. Compared with the inactivated EV71, the VP1 protein elicited similar humoral and cellular responses, but the engineered protein is safer, less expensive and can be produced more efficiently. Therefore, EV71‐VP1 protein can induce effective immunologic protection against EV71 and is an ideal candidate against EV71 infection.J. Med. Virol. 87:256–262, 2015.© 2014 Wiley Periodicals, Inc.