A scalable and reproducible manufacturing process for Phlebotomus papatasi salivary protein PpSP15, a vaccine candidate for leishmaniasis

Cutaneous leishmaniasis is a parasitic and neglected tropical disease transmitted by the bites of sandflies. The emergence cutaneous in areas war, conflict, political instability, climate change has prompted efforts to develop preventive vaccine. One vaccine candidate antigen PpSP15, 15 kDa salivary from sandfly Phlebotomus papatasi that facilitates infection Leishmania parasite been shown induce parasite-specific cell-mediated immunity. Previously, we developed fermentation process for producing recombinant PpSP15 Pichia pastoris two-chromatographic-step purification at 100 mL scale. Here expand design 10 L scale examine its reproducibility performing three identical runs, an essential transition step towards technology transfer pilot manufacture. was able reproducibly recover 81% protein with yield 0.75 g/L supernatant, purity level 97% low variance among runs. Additionally, freeze-thaw stability study indicated remains stable after undergoing cycles, accelerated confirmed 37 °C least one month. A research cell bank expression generated fully characterized. Collectively, production are ready future cGMP manufacturing.