Formulating mRNA-Loaded Nanoparticles for Vaccination Purposes
Published: May 31, 2024
Abstract
Source: Fornaguera, C. et al., Synthesis and Characterization of mRNA-Loaded Poly(Beta Aminoesters) Nanoparticles for Vaccination Purposes. J. Vis. Exp. (2021)
This video demonstrates a method for generating an mRNA vaccine candidate using polymeric nanoparticles. The cationic polymer interacts with negatively charged mRNA, forming polyplexes. The formation of polyplexes stabilizes the mRNA, making it a potential candidate for future mRNA vaccines in immunotherapeutic applications.
Protocol
1. Synthesis of poly(beta amino esters) (pBAE) polymer with end oligopeptides (OM-pBAE) Polymerization of C6-pBAE Add 5-amino-1-pentanol (38 mmol; MW = 103.16 Da) 1-hexylamine (38 mmol; MW = 101.19 Da) into a round-bottom glass flask (100 mL). Then, add 1,4-butanediol diacrylate (82 mmol; MW = 198.22 Da). Pre-heat the silicone oil bath at 90 °C, place the round-bottom flask into the oil bath and stir the mixture with the aid of a magnetic stir bar overnight (~…
Disclosures
The authors have nothing to disclose.
Materials
| 1,4-butanediol diacrylate | Sigma Aldrich | 123048 | |
| 1-hexylamine | Sigma Aldrich | 219703 | |
| 5-amino-1-pentanol | Sigma Aldrich | 411744 | |
| Acetone | Panreac | 141007 | |
| Chlor hydroxhyde | Panreac | 181023 | |
| Chloroform-d | Sigma Aldrich | 151823 | |
| Cys-His-His-His peptide | Ontores | Custom | |
| Cys-Lys-Lys-Lys peptide | Ontores | Custom | |
| D2O | Sigma Aldrich | 151882 | |
| DEPC reagent for Rnase free water | Sigma Aldrich | D5758 | This reagent is important to treat MilliQ water to remove any RNases of the buffers |
| Diethyl ether | Panreac | 212770 | |
| Dimethyl sulfoxide | Sigma Aldrich | 276855 | |
| HEPES | Sigma Aldrich | H3375 | |
| mRNA EGFP | TriLink Technologies | L-7601 | |
| RiboGreen kit | ThermoFisher | R11490 | |
| Sodium acetate | Sigma Aldrich | 71196 | |
| Sucrose | Sigma Aldrich | S0389 | |
| Trifluoroacetic acid | Sigma Aldrich | 302031 |
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Cite This Article
Formulating mRNA-Loaded Nanoparticles for Vaccination Purposes. J. Vis. Exp. (Pending Publication), e22266, doi: (2024).