High-Sensitivity RNA Quantification for LNP Encapsulation Efficiency

June 30, 2026

BlueskyReddit
University of Florida
Neurosurgery
Research Scientist

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Ease-of-Optimization

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Company:

Thermo Fisher Scientific (Invitrogen)

Product Name:

Quant-iT™ RiboGreen RNA Reagent

Catalog Number:

R11490

I used the Quant-iT™ RiboGreen High Sensitivity assay to accurately quantify encapsulated and unencapsulated mRNA in lipid nanoparticles (LNPs) for encapsulation efficiency measurements. The assay was highly reproducible, sensitive, and easy to optimize, making it ideal for formulation development and quality control.

Experimental Design and Results Summary

Application

Quantification of mRNA encapsulation efficiency in lipid nanoparticles (LNPs) using the Quant-iT™ RiboGreen High Sensitivity assay

Starting Material

Tips

Dilute LNP samples 1:200 in TE buffer and prepare a fresh High Sensitivity standard curve for each assay. Incubate samples with RiboGreen for 4 minutes before reading fluorescence (Ex 480 nm/Em 520 nm). For total RNA, incubate LNPs with 2.5% Triton X-100 at 37°C for 15 minutes before adding RiboGreen. Protect the reagent from light and use the same incubation time for all samples and standards

Results Summary

The assay accurately distinguished free from encapsulated mRNA by comparing untreated LNPs with Triton X-100-treated samples. A highly linear standard curve enabled reliable quantification of RNA over the working range, and encapsulation efficiency measurements were highly reproducible across independent experiments. The assay was sufficiently sensitive for routine characterization of mRNA-LNP formulations during formulation optimization and stability studies.

DOI or PMID #

N/A

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Summary

The Good

Excellent sensitivity for low RNA concentrations.

The Bad

The reagent is light-sensitive and requires careful handling.

The Bottom Line

The Quant-iT™ RiboGreen High Sensitivity assay has become our standard method for measuring mRNA encapsulation efficiency in lipid nanoparticles. The protocol is easy to implement, highly reproducible, and provides sensitive, accurate RNA quantification with minimal sample consumption. Combined with Triton X-100 lysis, it enables reliable determination of both free and total mRNA, making it an excellent tool for formulation development, optimization, and quality control of mRNA-LNP systems.

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