Evolution of the structure of lipid nanoparticles for nucleic acid delivery : From in situ studies of formulation to colloidal stability
(2024) In Journal of Colloid and Interface Science 660. p.66-76- Abstract
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)... (More)
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.
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- author
- Gilbert, Jennifer LU ; Sebastiani, Federica LU ; Arteta, Marianna Yanez ; Terry, Ann LU ; Fornell, Anna LU ; Russell, Robert ; Mahmoudi, Najet and Nylander, Tommy LU
- organization
- publishing date
- 2024-04-15
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Colloid and Interface Science
- volume
- 660
- pages
- 11 pages
- publisher
- Elsevier
- external identifiers
-
- pmid:38241872
- scopus:85182743795
- ISSN
- 0021-9797
- DOI
- 10.1016/j.jcis.2023.12.165
- language
- English
- LU publication?
- yes
- id
- 156111b2-6bef-4c7d-bb04-137418227fea
- date added to LUP
- 2024-02-16 14:26:19
- date last changed
- 2024-11-15 06:29:42
@article{156111b2-6bef-4c7d-bb04-137418227fea, abstract = {{<p>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.</p>}}, author = {{Gilbert, Jennifer and Sebastiani, Federica and Arteta, Marianna Yanez and Terry, Ann and Fornell, Anna and Russell, Robert and Mahmoudi, Najet and Nylander, Tommy}}, issn = {{0021-9797}}, language = {{eng}}, month = {{04}}, pages = {{66--76}}, publisher = {{Elsevier}}, series = {{Journal of Colloid and Interface Science}}, title = {{Evolution of the structure of lipid nanoparticles for nucleic acid delivery : From in situ studies of formulation to colloidal stability}}, url = {{http://dx.doi.org/10.1016/j.jcis.2023.12.165}}, doi = {{10.1016/j.jcis.2023.12.165}}, volume = {{660}}, year = {{2024}}, }