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 (~…

Divulgaciones

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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Formulating mRNA-Loaded Nanoparticles for Vaccination Purposes. J. Vis. Exp. (Pending Publication), e22266, doi: (2024).

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