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bioRxiv · 10.1101/2024.05.17.594777

Fabrication of mRNA encapsulated Lipid nanoparticles using State-of-the-art SMART-MaGIC Technology and transfection in vitro

Heshmati, N.·Chakka, L. R. J.·Zhang, Y.·Maniruzzaman, M.
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Abstract

The messenger ribose nucleic acid (mRNA) in lieu of Corona virus of 2019 (COVID-19) vaccines were effectively delivered through Lipid nanoparticles (LNP) which were proved effective carriers for clinical applications. In the present work, mRNA (erythropoietin (EPO)) encapsulated LNPs were prepared using a next generation state-of-the-art patented Sprayed Multi Absorbed-droplet Reposing Technology (SMART) coupled with Multi-channeled and Guided Inner-Controlling printheads (MaGIC) technologies. The LNP-mRNA were synthesized at different N/P ratios and the particles were characterized for particle size & zeta potential (Zetasizer), encapsulation or complexation (gel retardation assay) and transfection (Fluorescence microscopy) in MG63 sarcoma cells in vitro. The results showed a narrow distribution of mRNA-lipid particles of 200 nm when fabricated with SMART alone and then the size was reduced to approximately 50 nm with the combination of SMART-MaGIC technologies. The gel retardation assay showed that the N/P>1 showed strong encapsulation of mRNA with lipid. The in vitro results showed the toxicity profile of the lipids where N/P ratio of 5 is the optimized with >50% cell viability. The functional LNP-mRNA were prepared and analyzed with SMART-MaGIC technologies which could be a potential new fabrication method of mRNA loaded LNPs. HighlightsO_LIThe particle size was reduced to around 50 nm with implementation of SMART-MaGIC. C_LIO_LIThe loading efficiency is 100%. C_LIO_LIThe functionality of the mRNA is unaffected during the preparation process. C_LIO_LIThe transfection facilitated transient expression of the protein in vitro. C_LIO_LIThe EPO mRNA is more effective than EPO protein to reduce chemo-toxic effects in vitro. C_LI

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Heshmati, N., Chakka, L. R. J., Zhang, Y., Maniruzzaman, M.. 2024-05-21. Fabrication of mRNA encapsulated Lipid nanoparticles using State-of-the-art SMART-MaGIC Technology and transfection in vitro. https://doi.org/10.1101/2024.05.17.594777

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