Study on molecular orientation and stratification in RNA-lipid nanoparticles by cryogenic orbitrap secondary ion mass spectrometry
Name
s42004-025-01526-x.pdf
Description
Published version
Size
1.38 MB
Format
Adobe PDF
Checksum (MD5)
0cb85a902b78ffa51821dc9d8e0aa2e5
Author(s)
Kotowska, Anna M
Fay, Michael
Watts, Julie A
Gilmore, Ian S
Scurr, David J
Howe, Alaina
Capka, Vladimir
Perez, Corey E
Doud, Devin
Patel, Siddharth
Date Issued
May 22, 2025
Journal
Communications Chemistry
Publisher
Springer Science and Business Media LLC
Citation
Kotowska, A.M., Fay, M., Watts, J.A. et al. Study on molecular orientation and stratification in RNA-lipid nanoparticles by cryogenic orbitrap secondary ion mass spectrometry. Commun Chem 8, 160 (2025).
Version
Final published version
Abstract
Lipid nanoparticle RNA (LNP-RNA) formulations are used for the delivery of vaccines and other therapies. RNA molecules are encapsulated within their interior through electrostatic interactions with positively charged lipids. The identity of the lipids that present at their surface play a role in how they interact with and are perceived by the body and their resultant potency. Here, we use a model formulation to develop cryogenic sample preparation for molecular depth profiling Orbitrap secondary ion mass spectrometry (Cryo-OrbiSIMS) preceded by morphological characterisation using cryogenic transmission electron microscopy (Cryo-TEM). It is found that the depth distribution of individual lipid components is revealed relative to the surface and the RNA cargo defining the core. A preferential lipid orientation can be determined for the 1,2-Dimyristoyl-glycero-3-methox-polyethylene glycol 2000 (DMG-PEG2k) molecule, by comparing the profiles of PEG to DMG fragments. PEG fragments are found immediately during analysis of the LNP surface, while the DMG fragments are deeper, coincident with RNA ions located in the core, in agreement with established models of LNPs. This laboratory-based de novo analysis technique requires no labelling, providing advantages over large facility neutron scattering characterisation.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Koch Institute for Integrative Cancer Research at MIT
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DOI of Published Version
https://doi.org/10.1038/s42004-025-01526-x
