Formulating mRNA-Loaded Nanoparticles for Vaccination Purposes

Begin with a frozen polymer solution containing positively charged polymers.

Thaw the polymer solution until it reaches room temperature.

Add a buffer and vortex for uniform mixing, generating a homogeneous solution.

Introduce target mRNA to the polymer solution in an equal ratio for optimum reaction.

Incubate to allow the positively charged polymers to interact with the negatively charged phosphate groups in the mRNA via electrostatic interactions.

This facilitates the incorporation of the mRNA within the polymer structure, forming nanoparticles of mRNA-polymer complexes or polyplexes.

Transfer these polyplexes to a tube containing RNase-free water.

Mix them and allow the precipitation of the polyplexes.

Introduce a buffer containing sucrose and freeze it. Sucrose acts as a cryoprotectant that maintains the structural integrity of the polyplexes.

Then, perform freeze-drying, to remove any traces of water.

Store it at a low temperature as an mRNA vaccine candidate for future use.