Evolution of the structure of lipid nanoparticles for nucleic acid delivery: From in situ studies of formulation to colloidal stability

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  • Jennifer Gilbert
  • Sebastiani, Federica
  • Marianna Yanez Arteta
  • Ann Terry
  • Anna Fornell
  • Robert Russell
  • Najet Mahmoudi
  • Tommy Nylander

The development of lipid nanoparticle (LNP) based therapeutics for delivery of RNA has triggered the advance of new strategies for formulation, such as high throughput microfluidics for precise mixing of components into well-defined particles. In this study, we have characterised the structure of LNPs throughout the formulation process using in situ small angle x-ray scattering in the microfluidic chip, then by sampling in the subsequent dialysis process. The final formulation was investigated with small angle x-ray (SAXS) and neutron (SANS) scattering, dynamic light scattering (DLS) and cryo-TEM. The effect on structure was investigated for LNPs with a benchmark lipid composition and containing different cargos: calf thymus DNA (DNA) and two model mRNAs, polyadenylic acid (polyA) and polyuridylic acid (polyU). The LNP structure evolved during mixing in the microfluidic channel, however was only fully developed during the dialysis. The colloidal stability of the final formulation was affected by the type of incorporated nucleic acids (NAs) and decreased with the degree of base-pairing, as polyU induced extensive particle aggregation. The main NA LNP peak in the SAXS data for the final formulation were similar, with the repeat distance increasing from polyU<polyA<DNA, following the expected extent of base-pairing.

OriginalsprogEngelsk
TidsskriftJournal of Colloid and Interface Science
Vol/bind660
Sider (fra-til)66-76
ISSN0021-9797
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
This work benefited from the use of the SasView application, originally developed under National Science Foundation, NSF , award DMR-0520547 . SasView contains code developed with funding from the European Union's Horizon 2020 research and innovation programme under the SINE2020 project, grant agreement No 654000 . The deuterated cholesterol used in this work was provided by the National Deuteration Facility, which is partly funded by the National Collaborative Research Infrastructure Strategy (NCRIS) – an Australian Government initiative. Research conducted at MAX IV, a Swedish national user facility, was supported by the Swedish Research Council under contract 2018-07152 , the Swedish Governmental Agency for Innovation Systems under contract 2018-04969 , and Formas under contract 2019-02496 . We acknowledge the use of the “AdaptoCell for MAX IV Laboratory users” platform developed under the Swedish Foundation for Strategic Research , grant ITM-0375 . The financial support from Swedish Research Council under contract 2017-06716 and 2020-05421 and from NanoLund is gratefully acknowledged. We thank the ISIS Neutron and Muon source (UK) for allocating beamtime on Sans2d (RB1920532). We also would like to acknowledge Professor Joachim Rädler for inspirational discussion.

Funding Information:
This work benefited from the use of the SasView application, originally developed under National Science Foundation, NSF, award DMR-0520547. SasView contains code developed with funding from the European Union's Horizon 2020 research and innovation programme under the SINE2020 project, grant agreement No 654000. The deuterated cholesterol used in this work was provided by the National Deuteration Facility, which is partly funded by the National Collaborative Research Infrastructure Strategy (NCRIS) – an Australian Government initiative. Research conducted at MAX IV, a Swedish national user facility, was supported by the Swedish Research Council under contract 2018-07152, the Swedish Governmental Agency for Innovation Systems under contract 2018-04969, and Formas under contract 2019-02496. We acknowledge the use of the “AdaptoCell for MAX IV Laboratory users” platform developed under the Swedish Foundation for Strategic Research, grant ITM-0375. The financial support from Swedish Research Council under contract 2017-06716 and 2020-05421 and from NanoLund is gratefully acknowledged. We thank the ISIS Neutron and Muon source (UK) for allocating beamtime on Sans2d (RB1920532). We also would like to acknowledge Professor Joachim Rädler for inspirational discussion.

Publisher Copyright:
© 2023 The Author(s)

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