| Abstract: |
Due to low immune system response lipid nanoparticles (LNPs) have developed into an attractive alternative to AAVs for the delivery of nucleic acids for gene therapy. They are most frequently used for the transfer of short interfering RNA (si-RNA) or messenger RNA (mRNA). While the first ones are often loaded with several thousand copies per particle, only a few mRNA copies are delivered per LNP. This renders the exact loading state an important parameter for the dosage of LNP-mRNA therapeutics. Here, we show how density variation via different H20/D20 ratios can help to determine the copy number of mRNA per lipid nanoparticle. Density contrast SV-AUC enabled us to measure the lipid/mRNA ratio within a size distribution of special LNPs which are biodegradable, so called DLP (degradable lipid nanoparticles). Multi-signal SV-AUC allowed the orthogonal confirmation of our findings. |
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