Diphtheria and tetanus toxoids and acellular pertussis (DTaP) vaccines were widely used since 1940s. The exceptional success of childhood vaccination is undisputed. However, the anti-diphtheria and tetanus antibody will decrease with the increase of age in human body. A boosting vaccine for tetanus and diphtheria in adult is recommended by WHO. Recombinant protein vaccine has the advantages of single component and high safety, which is one of the directions to develop boosting vaccines. Therefore, in this study, we evaluated a recombinant TTc and CRM197 combination vaccine (RTCV) that uses the fragment C (TTc) of tetanus toxin and the cross-reacting material 197 (CRM197) of the diphtheria toxin mutant. Our results displayed that RTCV (composed of 10 mu g/mL TTc, 20 mu g/mL CRM197 antigens, and 500 mu g/mL aluminum adjuvants) could induce high levels of IgG and IgG1 antibody in mice, which were similar as those induced by DTaP. These results will provide technical support for a novel boosting vaccine against diphtheria and tetanus.