Lipid nanoparticles (LNPs) are effective vehicles to deliver mRNA vaccines and therapeutics. It has been challenging to assess mRNA packaging characteristics in LNPs, including payload distribution and capacity, which are critical to understanding structure-property-function relationships for further carrier development. Here, we report a method based on the multi-laser cylindrical illumination confocal spectroscopy (CICS) technique to examine mRNA and lipid contents in LNP formulations at the single-nanoparticle level. By differentiating unencapsulated mRNAs, empty LNPs and mRNA-loaded LNPs via coincidence analysis of fluorescent tags on different LNP components, and quantitatively resolving single-mRNA fluorescence, we reveal that a commonly referenced benchmark formulation using DLin-MC3 as the ionizable lipid contains mostly 2 mRNAs per loaded LNP with a presence of 40%-80% empty LNPs depending on the assembly conditions. Systematic analysis of different formulations with control variables reveals a kinetically controlled assembly mechanism that governs the payload distribution and capacity in LNPs. These results form the foundation for a holistic understanding of the molecular assembly of mRNA LNPs.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9508184 | PMC |
| http://dx.doi.org/10.1038/s41467-022-33157-4 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Immunoconjugates as an Efficient Platform for Drug Delivery: A Resurgence of Natural Products in Targeted Antitumor Therapy.
Pharmaceuticals (Basel)
December 2024
Department "Pharmacology, Pharmacotherapy and Toxicology", Faculty of Pharmacy, Medical University of Sofia, 1000 Sofia, Bulgaria.
The present review provides a detailed and comprehensive discussion on antibody-drug conjugates (ADCs) as an evolving new modality in the current therapeutic landscape of malignant diseases. The principle concepts of targeted delivery of highly toxic agents forsaken as stand-alone drugs are examined in detail, along with the biochemical and technological tools for their successful implementation. An extensive analysis of ADCs' major components is conducted in parallel with their function and impact on the stability, efficacy, safety, and resistance profiles of the immunoconjugates.
View Article and Find Full Text PDFImproving agricultural spraying with multi-rotor drones: a technical study on operational parameter optimization.
Front Nutr
December 2024
Directorate of Crop Management, Tamil Nadu Agricultural University, Coimbatore, India.
Article Synopsis
- Drones are improving nutrient management by allowing precise spraying techniques, which are especially important under climate change.
- Current research focuses on optimizing various parameters like flight height, pressure, and nozzle configuration to enhance spray efficiency and reduce waste.
- The study found that a hover height of 2.0 meters provided the best spray uniformity and distribution, showing the effectiveness of careful drone operation in agricultural practices.
Understanding the self-assembly and molecular structure of mRNA lipid nanoparticles at real size: Insights from the ultra-large-scale simulation.
Int J Pharm
December 2024
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau 999078, China; Faculty of Health Sciences, University of Macau, Macau 999078, China. Electronic address:
Messenger RNA (mRNA) encapsulated in lipid nanoparticles (LNPs) represents a cutting-edge delivery technology that played a pivotal role during the COVID-19 pandemic and in advancing vaccine development. However, molecular structure of mRNA-LNPs at real size remains poorly understood, with conflicting results from various experimental studies. In this study, we aim to explore the assembly process and structural characteristics of mRNA-LNPs at realistic sizes using coarse-grained molecular dynamic simulations.
View Article and Find Full Text PDFKinetic Modeling of the Antibody Disulfide Bond Reduction Reaction With Integrated Prediction of the Drug Load Profile for Cysteine-Conjugated ADCs.
Biotechnol Bioeng
December 2024
Institute of Process Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), Karlsruhe, Baden-Württemberg, Germany.
Antibody-drug conjugates (ADC) constitute a groundbreaking advancement in the field of targeted therapy. In the widely utilized cysteine conjugation, the cytotoxic payload is attached to reduced interchain disulfides which involves a reduction of the native monoclonal antibody (mAb). This reaction needs to be thoroughly understood and controlled as it influences the critical quality attributes (CQAs) of the final ADC product, such as the drug-to-antibody ratio (DAR) and the drug load distribution (DLD).
View Article and Find Full Text PDFModular functionalization of cellular nanodiscs enables targeted delivery of chemotherapeutics into tumors.
J Control Release
December 2024
Aiiso Yufeng Li Family Department of Chemical and Nano Engineering, Shu and K.C. Chien and Peter Farrell Collaboratory, University of California San Diego, La Jolla, CA 92093, USA. Electronic address:
The effective delivery of chemotherapeutic drugs to tumor sites is critical for cancer treatment and remains a significant challenge. The advent of nanomedicine has provided additional avenues for altering the in vivo distribution of drug payloads and increasing tumor localization. More recently, cell-derived nanoparticles, with their biocompatibility and unique biointerfacing properties, have demonstrated considerable utility for drug delivery applications.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!