Morphological changes induced by buffer exchange during preparation of mRNA-lipid nanoparticles occur in a stepwise manner as pH is elevated

Jan 19, 2026Molecular therapy. Methods & clinical development

Step-by-step shape changes in mRNA-lipid nanoparticles during buffer exchange as pH increases

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Abstract

Stable lipid nanoparticles (LNPs) with bleb-like protrusions were formed at an intermediate pH of 5.5.

  • Initial nucleic acid entrapment in LNPs relies on ionizable cationic lipids at low pH with ethanol.
  • FRET and cryo-electron microscopy confirmed that interactions and fusion of particles occurred during buffer exchange.
  • Different ionizable lipids resulted in varying particle behaviors, with SM-102 LNPs showing fusion and growth at higher pH values.
  • Small-angle X-ray scattering indicated that MC3 and ALC-0315 LNPs lost bilayer-like structures at a final pH of 7.4.
  • LNPs made with DODMA and SM-102 retained some bilayer-like structures at pH 7.4.

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