Preparation of a Nanoemulsion Adjuvant Vaccine

Abstract

Source: Zhang, Z. et al., Preparation, Characteristics, Toxicity, and Efficacy Evaluation of the Nasal Self-Assembled Nanoemulsion Tumor Vaccine In Vitro and In Vivo. J. Vis. Exp. (2022)

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

1. 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.
2. Add Tween 80 and IKVAV-OVA_257-264 (I-OVA) quantitatively and mix.         
   NOTE: Tween 80 and I-OVA were mixed at a mass ratio of 25:1.
3. Add squalene into the mixture prepared in step 1.2 (7:3, Smix: squalene).
4. Add 100 µL of MPLA solution (10 mg/mL) to the mixture prepared in step 1.3.
5. 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.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
MPLA	Invivogen Lit. Co.	tlrl-mpla
OVA257–264	Shanghai Botai Biotechnology Co., Ltd.	NA
Synthetic peptide (I-OVA) conjugation of IKVAV-PA	Shanghai Botai Biotechnology Co., Ltd.	NA