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mRNA has shown therapeutic potential in a range of applications, including viral vaccines, protein replacement therapies, cancer immunotherapies, cellular reprogramming and genome editing. To achieve therapeutic effects, mRNA molecules have to reach specific target cells and produce sufficient proteins of interest. Lipid nanoparticles (LNPs) are the leading nonviral technologies for the delivery of exogenous RNA to target cells in vivo. A major challenge to advance lipid nanoparticles (LNPs) for RNA therapeutics is the development of formulations that can be produced reliably across the various scales of drug development.
Seattle Genova is highly specialized in synthesizing top-quality mRNA and lipid nanoparticles for research and development applications, focusing on tailored production to meet your specific needs. We also offer a standard portfolio with optimized stability, solubility, and handling characteristics.
LNPs are typically composed of glycerophospholipids, canonic lipids, sterol lipids, and PEGylated lipids that encapsulate oligonucleotides, which are contained within an aqueous phase.
Table 1: Commonly used mRNA delivery systems
Lipid Nanoparticle | Liposome | Solid Lipid Nanoparticle | Nanostructured Lipid Carrier | |
Lipid Shell | Monolayer | Bilayer | Surfactant | Surfactant |
Internal Core | Reverse micelles | Aqueous | Solid lipids | Solid and liquid lipids |
Cargo | Oligonucleotides | Hydrophobic and/or hydrophilic small molecules | Hydrophobic and/or hydrophilic small molecules | Hydrophobic and/or hydrophilic small molecules |
Size | ~20-100 nm | ~50-1,000 nm | ~40-1,000 nm | ~40-1,000 nm |
Generally, mRNA-LNP design is comprised of several elements: (1) mRNA sequence design and nucleotide modification choice, (2) Optimization of a LNP formulation to encapsulate and deliver mRNA and (3) long-term storage. Each one of these phases entails a plethora of choices and considerations to make.
Workflow of mRNA Preparation:
1.Antigen design
2.Plasmid production
3.In vitro transcription
4.Characterization
A, primary structure of mRNA
-Sequencing
-Mapping
-mRNA modification analysis
B, mRNA secondary structure
-CD
-Cryo-EM
-Secondary structure prediction
C, mRNA product analysis
-mRNA integrity and purity analysis
-analysis of mRNA capping efficiency
-Poly A tail length analysis
-Double-Stranded RNA Analysis
Procedures for the preparation of LNPs:
1.Design
-Lipid molar ratio
-Lipid:oligonucleotide weight ratio
-Ionizable lipid nitrogen:oligonucleotide phosphate (N:P) molar ratio
-Lipid acid dissociation constant (lipid pK)
-Buffer composition, ionic strength, and pH
-Particle size and pharmacokinetics
-Routes of administration for LNPs
2.Preparation and mixing
-Mixing
-Extrusion
-Dialysis
-Filter-sterilize
3.Stability
-Physical stability
-Chemical stability
-Biological stability
4.Storage
-Short-term storage at 4°C
Thank you for your interest in our LNP Formulated RNA Production Service services. Please complete the form below and we will contact you shortly.
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