Recombinant antigens and adjuvant
The recombinant TSA-1-C4 and Tc24-C4 antigens were obtained from the Departamento de Biotecnología y Bioingeniería from the Centro de Investigación y Estudios Avanzados (CINVESTAV) of the Instituto Politécnico Nacional (IPN), México. Each TSA-1-C4 or Tc24-C4 coding sequence was cloned into a pET41a+ E. coli expression vector26,30. The resulting plasmid DNA was transformed into BL21 (DE3) cells induced with isopropyl-beta-D-1-thiogalactoside (IPTG) for protein expression. Recombinant proteins were purified by ion exchange (IEX) and size exclusion chromatography (SEC)30. The integrity and size of each recombinant protein was analysed by SDS-PAGE electrophoresis (Supplementary Fig. 1 ). For cytotoxicity assay, the recombinant proteins were formulated with the adjuvant E6020 in a stable squalene emulsion (SE). E6020 Toll-like receptor 4 agonist was acquired through Eisai, Inc34.