Solution biophysics identifies lipid nanoparticle non-sphericity, polydispersity, and dependence on internal ordering for efficacious mRNA delivery DOI Creative Commons
Marshall S. Padilla, Sarah J. Shepherd, Andrew R. Hanna

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 22, 2024

Abstract Lipid nanoparticles (LNPs) are the most advanced delivery system currently available for RNA therapeutics. Their development has accelerated since success of Patisiran, first siRNA-LNP therapeutic, and mRNA vaccines that emerged during COVID-19 pandemic. Designing LNPs with specific targeting, high potency, minimal side effects is crucial their successful clinical use. These characteristics have been improved through microfluidic platforms, which enhanced efficacy uniformity LNP batches. However, our understanding how composition mixing method influences structural, biophysical, biological properties resulting particles remains limited, hindering LNPs. Our lack structural extends from physical compositional polydispersity LNPs, render traditional characterization methods, such as dynamic light scattering (DLS), unable to accurately quantitate physicochemical In this study, we address challenge structurally characterizing polydisperse formulations using emerging solution-based biophysical methods higher resolution provide data beyond size polydispersity. techniques include sedimentation velocity analytical ultracentrifugation (SV-AUC), field flow fractionation followed by multi-angle (FFF-MALS), size-exclusion chromatography in-line synchrotron small-angle X-ray (SEC-SAXS). Here, show intrinsic in size, loading, shape, these parameters dependent on both formulation technique lipid composition. Lastly, demonstrate can be employed predict transfection three models examining relationship between translation characteristics. We envision employing will essential determining structure-function relationships, facilitating creation new design rules future

Language: Английский

Computational Methods for Modeling Lipid-Mediated Active Pharmaceutical Ingredient Delivery DOI Creative Commons
Markéta Paloncýová, Mariana Valério, Ricardo Nascimento dos Santos

et al.

Molecular Pharmaceutics, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 29, 2025

Lipid-mediated delivery of active pharmaceutical ingredients (API) opened new possibilities in advanced therapies. By encapsulating an API into a lipid nanocarrier (LNC), one can safely deliver APIs not soluble water, those with otherwise strong adverse effects, or very fragile ones such as nucleic acids. However, for the rational design LNCs, detailed understanding composition-structure-function relationships is missing. This review presents currently available computational methods LNC investigation, screening, and design. The state-of-the-art physics-based approaches are described, focus on molecular dynamics simulations all-atom coarse-grained resolution. Their strengths weaknesses discussed, highlighting aspects necessary obtaining reliable results simulations. Furthermore, machine learning, i.e., data-based approach to lipid-mediated introduced. data produced by experimental theoretical provide valuable insights. Processing these help optimize LNCs better performance. In final section this Review, computer reviewed, specifically addressing compatibility

Language: Английский

Citations

2
Rheology of mRNA-loaded lipid nanodumbbells DOI
M. A. Kanso, Shalini Singh, A. Jeffrey Giacomin

et al.

Physics of Fluids, Journal Year: 2025, Volume and Issue: 37(2)

Published: Feb. 1, 2025

In one important chemical engineering unit operation of messenger ribonucleic acid (mRNA) vaccine manufacture, the precious mRNA payload is encapsulated in lipid nanoparticles (LNPs). Recent elegant cryogenic-transmission electron microscopy [Brader et al., Biophys. J. 120, 2766 (2021)] reveals that these take form dumbbell suspensions. When encapsulating their payloads, dumbbells can be both lopsided and interpenetrating, with smaller two beads carrying payload. this work, we arrive at analytical expressions for suspensions nanoparticle payloads. For this, first exploit rigid theory [Abdel-Khalik Bird, Appl. Sci. Res. 30, 268 (1975)], which relies on orientation distributions dumbbells, to predict suspension rheology, specifically how departs from Newtonian behavior. We next elastic [Phan-Thien Phys. Fluids 36, 071707 (2024)], also add stretching. Our results include relaxation time, rotational diffusivity, zero-shear viscosity, shear stress function, steady-shear viscosity viscous part minus complex viscosity. determine diffusivity mRNA-loaded nanodumbbells small-amplitude oscillatory measurements.

Language: Английский

Citations

1
Formulation screening of lyophilized mRNA-lipid nanoparticles DOI Creative Commons
Anna Ruppl,

Denis Kiesewetter,

Monika Koell-Weber

et al.

International Journal of Pharmaceutics, Journal Year: 2025, Volume and Issue: unknown, P. 125272 - 125272

Published: Jan. 1, 2025

Language: Английский

Citations

0
Investigating the stability of RNA-lipid nanoparticles in biological fluids: Unveiling its crucial role for understanding LNP performance DOI Creative Commons
Heyang Zhang, Matthias Barz

Journal of Controlled Release, Journal Year: 2025, Volume and Issue: 381, P. 113559 - 113559

Published: Feb. 27, 2025

Language: Английский

Citations

0
Lyophilized monkeypox mRNA lipid nanoparticle vaccines with long-term stability and robust immune responses in mice DOI Creative Commons
Bin Wang,

Quanyi Yin,

Yi Li

et al.

Human Vaccines & Immunotherapeutics, Journal Year: 2025, Volume and Issue: 21(1)

Published: March 11, 2025

The World Health Organization (WHO) has recently declared another global health emergency due to the rapidly spreading monkeypox (Mpox) outbreak in numerous African countries. To address unmet need contain using existing vaccines, this study developed a lyophilization process for an effective, scalable and affordable Mpox mRNA-LNP vaccine candidate crisis. A comprehensive evaluation optimization of formulation (the type/concentration cryoprotectants, buffer system, as well mRNA concentration reconstitution solvent) freeze-drying parameters (freezing method, temperature, cooling rate primary/secondary drying conditions) were conducted. freeze-dried product exhibits uniform appearance moisture content less than 1%. Reconstitution lyophilized resulted equivalent particle size/polydispersity index, encapsulation efficiency integrity compared that freshly prepared mRNA-LNP. Furthermore, can be scaled up 100-fold 2000 vials/batch. Notably, demonstrated storage stability at least 12 months 4°C, ambient temperature minimum 8 h post-reconstitution, exhibiting minimal deterioration quality. vitro biological activity vivo immunogenicity was comparable These results provide compelling rationale utilization technology enhancing accessibility developing countries, strategy is crucial containing epidemic infection.

Language: Английский

Citations

0
Process development of tangential flow filtration and sterile filtration for manufacturing of mRNA-lipid nanoparticles: A study on membrane performance and filtration modeling DOI
Wenjun Wu, Líliam Teixeira Oliveira,

A. Jain

et al.

International Journal of Pharmaceutics, Journal Year: 2025, Volume and Issue: unknown, P. 125520 - 125520

Published: March 1, 2025

Language: Английский

Citations

0
A Polysorbate-Based Lipid Nanoparticle Vaccine Formulation Induces In Vivo Immune Response Against SARS-CoV-2 DOI Creative Commons

Aishwarya Saraswat,

Alireza Nomani,

Lin-Kin Yong

et al.

Pharmaceutics, Journal Year: 2025, Volume and Issue: 17(4), P. 441 - 441

Published: March 29, 2025

Background: Lipid nanoparticles (LNPs) have proven effective in delivering RNA-based modalities. Rapid approval of the COVID-19 vaccines highlights promise LNPs as a delivery platform for nucleic acid-based therapies and vaccines. Nevertheless, improved LNP designs are needed to advance next-generation other gene toward greater clinical success. components formulation excipients play central role biodistribution, immunogenicity, stability. Therefore, it is important understand, identify, assess appropriate lipid developing safe formulation. Herein, this study focused on novel Polysorbate-80 (PS-80)-based LNP. We hypothesized that substituting conventional linear PEG-lipids with PS-80, widely used, biocompatible injectable surfactant featuring branched PEG-like structure, may change biodistribution pattern enhance long-term By leveraging PS-80’s unique structural properties, aimed develop an mRNA-LNP extrahepatic robust freeze/thaw tolerance. Methods: employed stepwise optimization establish both composition buffer yield stable, high-performing PS-80-based SARS-CoV-2 (SC2-PS80 LNP). compared phosphate- versus tris-based buffers stability, examined multiple ratios, evaluated impact incorporating PS-80 (a PEG-lipid) vivo biodistribution. Various analytical assays were performed particle size, encapsulation efficiency, mRNA purity, vitro potency developed humanized mouse model was used measure its immunogenicity over six months storage at −80 °C. Results: Replacing standard 1,2-dimyristoyl-rac-glycero-3-methoxy polyethylene glycol-2000 (PEG-DMG) increased spleen-specific expression mRNA-LNPs after intramuscular injection. Formulating tris-sucrose-salt (TSS) preserved LNP’s physicochemical properties °C, whereas PBS-sucrose (PSS) less protective under frozen conditions. Notably, TSS-based SC2-PS80 elicited potent humoral immunity mice, including high anti-spike IgG titers pseudovirus neutralization, comparable freshly prepared formulations. Conclusions: A formulated TSS confers delivery, strong against following months. These findings offer promising pathway design therapeutics enhanced stability potential.

Language: Английский

Citations

0
In vivomRNA delivery to the lung vascular endothelium by dicationic Charge-Altering Releasable Transporters DOI Creative Commons

Mahmoud M. AbdElwakil,

Jun Ni, Summer Ramsay-Burrough

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: April 19, 2025

Abstract Endothelial cells (EC) comprise the pulmonary vascular bed and play a significant role in health disease. Consequently, EC niche represents an attractive therapeutic target for treating wide range of diseases. We have identified new class dicationic Charge-Altering Releasable Transporters. These single-component transporters selectively deliver mRNA to lung upon intravenous administration without use targeting ligand. Significantly, number spatial array cationic charges within repeating units CART polymer are found control both delivery efficacy tissue tropism. High-resolution imaging revealed efficient endothelial arteries, veins capillaries. The selective tropism these CARTs, coupled with tunable synthesis this family amphiphiles, enabling platform research clinical applications.

Language: Английский

Citations

0
Enhancing RNA encapsulation quantification in lipid nanoparticles: Sustainable alternatives to Triton X-100 in the RiboGreen assay DOI Creative Commons
David P. Schultz, Rasmus Münter,

Alex Masi

et al.

European Journal of Pharmaceutics and Biopharmaceutics, Journal Year: 2024, Volume and Issue: 205, P. 114571 - 114571

Published: Oct. 28, 2024

Language: Английский

Citations

2
Nano Plasma Membrane Vesicle‐Lipid Nanoparticle Hybrids for Enhanced Gene Delivery and Expression DOI Creative Commons
Claudio Luca Alter, Claudia Lotter, Ramya Deepthi Puligilla

et al.

Advanced Healthcare Materials, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 10, 2024

Lipid nanoparticles (LNPs) have emerged as the leading nonviral nucleic acid (NA) delivery system, gaining widespread attention for their use in COVID-19 vaccines. They are recognized efficient NA encapsulation, modifiability, and scalable production. However, LNPs face efficacy potency limitations due to suboptimal intracellular processing, with endosomal escape efficiencies (ESE) below 2.5%. Additionally, up 70% of NPs undergo recycling exocytosis after cellular uptake. In contrast, cell-derived vesicles offer biocompatibility high-delivery but challenging load exogenous NAs manufacture at large-scale. To leverage strengths both systems, a hybrid system is designed by combining vesicles, such nano plasma membrane (nPMVs), through microfluidic mixing subsequent dialysis. These hybrids demonstrate tenfold increase ESE an 18-fold rise reporter gene expression vitro vivo zebrafish larvae (ZFL) mice, compared traditional LNPs. improvements linked unique physico-chemical properties, composition, morphology. By incorporating this strategy streamlines development process, significantly enhancing systems without need extensive screening.

Language: Английский

Citations

2