Get the skinny on LNPs with Stunner

Importance of the topic

Dynamic light scattering (DLS) and RNA quantification are essential for lipid nanoparticle (LNP) characterization, underpinning the development of gene therapies and mRNA vaccines.

Objectives and overview of the study

This application note evaluates the Stunner platform, which integrates high throughput DLS and short pathlength UV/Vis absorbance to deliver rapid, comprehensive analysis of RNA-containing LNPs, including particle size, size distribution, total RNA concentration and turbidity, using minimal sample volume.

Methodology and used instrumentation

Stunner combines automated DLS measurements on up to 96 samples with deconvolution of UV/Vis spectra to separate contributions from RNA, turbidity and other components.
Fluc-mRNA-LNPs and empty LNPs provided by Precision NanoSystems were diluted in PBS (pH 7.4) and measured in a microvolume SBS plate format.
RNA quantification was based on absorbance at 260 nm using a 2 µL sample, six replicates and deconvolution algorithms.
DLS calculations used a viscosity of 1.002 mPa·s and refractive index of 1.334 at 20 °C.

Main results and discussion

• UV/Vis spectra were deconvoluted to quantify total RNA concentration without dyes or reagents, eliminating interference from turbidity.
• RNA measurements by Stunner closely matched RiboGreen assays, with linear response (R2 > 0.99) down to 1.2 µg/mL.
• High throughput DLS characterized 96 samples in under one hour, reporting a mean hydrodynamic diameter of 79 nm ± 1% and PDI of 0.14 ± 0.02.
• Turbidity correlated linearly with particle concentration, with slope dependent on particle size and composition.

Benefits and practical applications of the method

• Reagent-free and dilution-free RNA quantification reduces assay complexity, time and cost.
• Low sample volume (2 µL) preserves valuable material.
• High throughput screening accelerates R&D, QA/QC and formulation development workflows.
• Integrated platform simplifies data management and supports regulatory compliance (21 CFR Part 11, USP/EP).

Future trends and potential applications

Integration of Stunner into automated and continuous monitoring systems could enable real-time quality control in manufacturing.
Standardization of turbidity-based concentration measurements may extend its use to other nanoparticle formulations and payloads.
Enhanced deconvolution algorithms and broader spectral analysis could improve characterization of complex colloidal systems.

Conclusion

The Stunner platform delivers rapid, accurate and comprehensive analysis of RNA-LNPs by uniting high throughput DLS with UV/Vis deconvolution, streamlining workflows and supporting accelerated development of nucleic acid therapeutics.

References

1. Walsh C, others. RiboGreen assay protocol. Precision NanoSystems User Guide PNI-SOP-S9-001-EXT. 2016.
2. Roces C, others. Manufacturing considerations for the development of lipid nanoparticles using microfluidics. Pharmaceutics. 2020;12(11):1–19.
3. Ribeiro L, others. Use of nanoparticle concentration as a tool to understand the structural properties of colloids. Scientific Reports. 2018;8(1):1–8.