Formulation and Preparation of Adjuvanted Immunogens

Oil-in-Water formulated TLR-4 agonist (GLA-SE, EM048) was produced by the Infectious Disease Research Institute (IDRI) [26 (link),27 (link)]. EM048 was mixed with recombinant protein in PBS to a final concentration of 50 μg/ml EM048 and 200 μg/ml recombinant protein. Sigma adjuvant system (Sigma) was reconstituted according to the manufacturer’s instructions and combined with recombinant protein to a final concentration of 200 μg/ml recombinant protein. Imject Alum (Thermo Scientific) was mixed with recombinant protein according to the manufacturer’s instructions to a final volume ratio of Imject Alum to immunogen of 1:1 and a final concentration of 200 μg/ml recombinant protein.