Highly attenuated modified vaccinia Ankara(MVA) is sensitive to repeat freeze-thaw cycle and easy to lose activity. In order to make the activity of MVA vaccine remain stable during its manufacturing, storage, and administration, the lyophilization as a good option could be resorted to; through screening, the right stabilizer composition and its production procedure were obtained. The final moisture content of freezing-dried recombinant MVA-HIV vaccine was lower than 3%. It can be reconstituted quickly and shows regular physical appearance and stable potency. In vivo functional experiment, mice were divided randomly into the liquid vaccination group, the lyophilized vaccination group, and the control group. Having been DNA vaccine priming, the mice were boosted with a dose of 107 pfu MVA-HIV vaccine, which produced indistinguishable antibody titer and cytotoxic T-lymphocyte(CTL) level compared with those of liquid vaccination group(P>0.05). These results demonstrate that lyophilized MVA vaccine can induce high immunogenicity in mice. Highly attenuated modified vaccinia Ankara (MVA) is sensitive to repeat freeze-thaw cycle and easy to lose activity. In order to make the activity of MVA vaccine remain stable during its manufacturing, storage, and administration, the lyophilization as a good option could be The final moisture content of freezing-dried recombinant MVA-HIV vaccine was lower than 3%. It can be readily cured and its regular procedure appearance and stable potency. In vivo functional experiment, mice were divided randomly into the liquid vaccination group, the lyophilized vaccination group, and the control group. Having been DNA vaccine priming, the mice were boosted with a dose of 107 pfu MVA-HIV vaccine, which produced indistinguishable antibody titer and cytotoxic T-lymphocyte (CTL) level compared with those of liquid vaccination group (P> 0.05). These results demonstrate that lyophilized MV A vaccine can induce high immunogenicity in mice.