Formulating mRNA-Loaded Nanoparticles for Vaccination Purposes

For polyplexes formation, thaw the previously prepared polymers C-6 peptide poly-beta amino esters and vortex the solution. After pipetting the polymer mix up and down, prepare a 12.5-millimolar solution in sodium acetate. Vortex the solution and wait for 10 minutes.

Next, prepare mRNA at 0.5 milligrams per milliliter and mix by pipetting. Vortex the polymer solution again. Then, mix the genetic material solution in the polymer solution in a one-to-one ratio in a microcentrifuge tube, and incubate the tube at 25 degrees Celsius for 30 minutes in a thermal block. After incubation, precipitate the mixture with one to two RNase-free water by adding the sample to a microcentrifuge tube containing the water.

Then, to include the excipients, add 20-millimolar HEPES and 4 percent sucrose solution in the same volume as the mixture of mRNA and poly-beta amino esters and mix by pipetting. For lyophilization, after freezing the polyplex solution at minus 80 degrees Celsius for one hour, perform the primary drying, then immediately store the polyplexes at minus 20 degrees Celsius to avoid rehydration.