Region II of the erythrocyte-binding antigen (EBA-175 RII) has been identified as a promising target for a malaria vaccine. A systematic approach to identify optimal preformulation conditions of a non-glycosylated (NG) antigen, EBA-175 RII-NG, has been developed. This approach consists of development of an empirical temperature/pH phase diagram, high throughput stabilizer screening and aluminum salt adjuvant adsorption studies. Using these physical methods, we developed a stable formulation for EBA-175 RII-NG at pH 6.0 with sucrose and Brij 35 as stabilizers and Adju-Phos as an adjuvant. This approach should be generally applicable to guiding the development of stable vaccine formulations.