A label-free multi-parameter screening method for predicting the stability of mRNA-LNP formulations

Area of Science:

• Biotechnology
• Pharmaceutical Sciences
• Materials Science

Background:

• Messenger RNA (mRNA) technologies are advancing rapidly, with lipid nanoparticles (LNPs) becoming crucial for vaccine and gene therapy delivery.
• The colloidal stability of mRNA-LNP formulations directly impacts their therapeutic effectiveness.
• Current methods for predicting formulation stability are often slow and complex.

Purpose of the Study:

• To develop a rapid, label-free, real-time method for assessing mRNA-LNP colloidal stability.
• To introduce new thermal stability readouts for efficient formulation screening.
• To compare the developed method with conventional accelerated stability studies.

Main Methods:

• Established a multi-parameter thermal profiling assay monitoring turbidity, cumulant radius (r h), and scattering in real-time.
• Evaluated six mRNA-LNP formulations with variations in buffer, preservative, and pH.
• Introduced aggregation-onset temperature (Tagg) and change in r h (Δr h) as key stability indicators.

Main Results:

• The thermal profiling method successfully captured thermally induced colloidal stability changes in mRNA-LNPs.
• A redefined aggregation-onset temperature (Tagg) and Δr h effectively identified early structural perturbations.
• Logistic regression modeling using Tagg and Δr h showed good agreement with 40°C accelerated stability data.

Conclusions:

• The developed thermal profiling strategy provides a rapid and efficient tool for early-stage screening of mRNA-LNP formulations.
• This method significantly reduces evaluation time compared to conventional stability studies.
• Further validation is needed to confirm applicability across diverse mRNA cargos and lipid compositions.