Mechanism of macromolecular structure evolution in self-assembled lipid nanoparticles for siRNA delivery

Marian E. Gindy, Katherine Difelice, Varun Kumar, Robert K. Prud'Homme, Robert Celano, R. Matthew Haas, Jeffrey S. Smith, David Boardman

Research output: Contribution to journalArticlepeer-review

38 Scopus citations


Lipid nanoparticles (LNPs) are a leading platform for therapeutic delivery of small interfering RNAs (siRNAs). Optimization of LNPs as therapeutic products is enabled by the development of structure-activity relationships (SAR) linking LNP physiochemical and macromolecular properties to bioperformance. Methods by which LNP properties can be rationally manipulated are thus critical enablers of this fundamental knowledge build. In this work, we present a mechanistic study of LNP self-assembly via a rapid antisolvent precipitation process and identify critical physiochemical and kinetic parameters governing the evolution of LNP three-dimensional macromolecular structure as a biorelevant SAR feature. Using small-angle X-ray scattering, LNPs are shown to undergo a temporal evolution in macromolecular structure during self-assembly, rearranging from initially disordered phases after precipitation into well-ordered structures following a necessary annealing stage of the assembly sequence. The ability of LNPs to undergo structural reorganization is shown to be effected by the chemical nature of the aqueous antisolvent used for precipitation. Antisolvents of varying buffering species differentially influence LNP macromolecular features, revealing a new participatory role of buffer ions in LNP self-assembly. Furthermore, the formation of macromolecular structure in LNPs is shown to improve the efficiency of siRNA encapsulation, thereby offering a simple, nonchemical route for preparation of high-payload LNPs that minimize the dose of lipid excipients. The developed LNP precipitation process and mechanistic understanding of self-assembly are shown to be generalizable, enabling the production of LNPs with a tunable range of macromolecular features, as evidenced by the cubic, hexagonal, and oligo-lamellar phase LNPs exemplarily generated.

Original languageEnglish (US)
Pages (from-to)4613-4622
Number of pages10
Issue number16
StatePublished - Apr 29 2014

All Science Journal Classification (ASJC) codes

  • Condensed Matter Physics
  • General Materials Science
  • Spectroscopy
  • Surfaces and Interfaces
  • Electrochemistry


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