Encapsulation of mRNA in Therapeutics Like Lipid Nanoparticles Probed by Deep-UV Resonance Raman Spectroscopy

Area of Science:

• Biochemistry
• Spectroscopy
• Nanotechnology

Background:

• Messenger RNA (mRNA) therapeutics show promise but face challenges in stability and delivery.
• Efficient mRNA delivery often requires encapsulation within lipid nanoparticles (LNPs).
• Quantifying mRNA encapsulation is crucial for optimizing LNP-based therapeutics.

Purpose of the Study:

• To quantify free and encapsulated mRNA within LNPs using deep-UV resonance Raman (DUVRR) spectroscopy.
• To develop a spectroscopic method for analyzing mRNA-lipid interactions.
• To demonstrate the utility of DUVRR spectroscopy for complex biochemical systems.

Main Methods:

• Utilized deep-UV resonance Raman (DUVRR) spectroscopy with 266 nm excitation.
• Acquired Raman spectra from samples with varying lipid concentrations.
• Applied Principal Component Analysis (PCA) and two-trace two-dimensional correlation spectroscopy (2T2D-COS).

Main Results:

• DUVRR spectroscopy differentiated mRNA based on encapsulation levels.
• Identified spectral variations linked to lipid presence, particularly between 1270-1800 cm⁻¹.
• Established a 1322 cm⁻¹ band as a quantitative indicator of lipid-mRNA interaction using 2T2D-COS.

Conclusions:

• DUVRR spectroscopy is a powerful tool for analyzing mRNA within LNPs.
• 2T2D-COS provides a novel quantitative method for complex biochemical analysis.
• This technique has significant potential for optimizing mRNA therapeutic development.