This video demonstrates the production of nanoemulsion tumor vaccines. The stable oil reservoir depots carrying synthetic peptides and adjuvants are produced by low-energy nanoemulsion, offering significant promise for tumor immunotherapy.
Protocol
1. Preparation of the I-OVA NE
Admix 1 mg of monophosphoryl lipid A (MPLA) with 100 µL of dimethyl sulfoxide (DMSO), vortex for 5 min, and allow it to stand for 4 h at room temperature (RT) to dissolve completely.
Add Tween 80 and IKVAV-OVA257-264 (I-OVA) quantitatively and mix. NOTE: Tween 80 and I-OVA were mixed at a mass ratio of 25:1.
Add squalene into the mixture prepared in step 1.2 (7:3, Smix: squalene).
Add 100 µL of MPLA solution (10 mg/mL) to the mixture prepared in step 1.3.
Prepare the nanoemulsion vaccine using low-energy emulsification methods: add the mixed solution to water droplets at approximately 70% of the total volume and stir gently to obtain a transparent and easily flowing mixture. NOTE: The BNE control (blank emulsion) was prepared by the same method, replacing water with I-OVA.