Applications of lipid nanoparticles (LNPs) are limited by their accumulation in the liver. The absence of a targeted delivery plan of action usually leads to the hepatic tissue.
How can we deliver LNPs loaded with therapeutic molecules to specific tissues and organs?
Wang et al. offer a simple and convenient answer to this question in a recent article published in Nature Protocols.
Selective organ-targeting molecules for precise mRNA delivery
LNPs’ physicochemical similarity to very low-density lipoprotein and their ability to adsorb apolipoprotein E in blood plasma makes them targets for the low-density lipoprotein receptor, which is highly expressed in the liver. While this feature may benefit intravenously delivering therapeutic molecules to the hepatocytes, specifically mRNA, it limits LNPs’ full potential.
LNPs can be engineered to overcome this limitation by adding a selective organ-targeting (SORT) molecule to their composition. SORT molecules facilitate LNPs redirection and cellular uptake in target organs via an endogenous targeting mechanism.
Tune tissue tropism relies on the SORT molecule’s presence of distinct chemical functional groups and the physicochemical properties (permanently cationic, anionic, ionizable cationic, zwitterionic, etc.). This, added to the quantity of incorporated SORT molecule, could result in a manageable translation of mRNA to protein within a specific organ or across a range of organs.
DIVERSA’s lipid nanoemulsions can also cross this boundary. They can be engineered to target distinct organs and tissues precisely. Our lipid nanoemulsions can exhibit specialized targeting ligands or receptor-binding domains, encompassing proteins or peptides adept at engaging with cell-surface receptors or markers.
How are selective organ-targeting lipid nanoparticles prepared?
SORT molecules can be added to the traditional composition mixture of LNPs. Formation of LNPs occurs via the ethanol dilution method wherein the ethanol solution containing all the lipids is rapidly mixed with the aqueous buffer solution containing mRNA.
The processes for mixing are straightforward, convenient, and, in most cases, do not require specialized instrumentation.
The authors propose three different molecules to deliver LNPs to the liver, brain, or lung and three mixing methods:
- Pipette mixing: Pipette mixing is the most common method to prepare small-scale batches of LNPs. It involves simply mixing two solutions by rapidly pipetting up and down by hand for a specific time to allow the formation of nanoparticles.
- Vortex mixing: to prepare medium-scale batches of LNPs. It consists of rapidly mixing the two solutions under vigorous vortexing for a specific time to allow nanoparticle formation.
- Microfluidic mixing: to prepare medium to large-scale batches of LNPs. Homemade or commercial microfluidic devices can be used. This method allows more reproducibility, LNPs uniformity, and controllability of mixing parameters that can influence LNPs size. However, it is also more technical and expensive and is advised for manufacturing consistent batches for in vivo animal studies or potential clinical use where large scales are required.
The protocols do not require specific expertise, are modular to various lipids within defined physicochemical classes, and can be accomplished by researchers from different backgrounds.
DIVERSA: simple solutions for complex challenges
At DIVERSA, we have pioneered the development of highly adaptable lipid nanoemulsions designed to precisely deliver peptides, proteins, nucleic acids, and small molecules. Our technology is the best example to highlight the abovementioned methods since our nanoemulsions can be produced through the straightforward pipette mixing technique.
Simple methods such as pipette mixing obviate the need for specialized instrumentation, offering an adaptable and cost-effective technology. It can be executed in any laboratory, catering to scientists from various disciplines. This provides a future of unparalleled technological advancements in different therapeutic sectors with clearly unmet clinical needs. Spanning research in oncology, inflammatory and autoimmune diseases, and metabolic disorders, it also extends its relevance to nutraceuticals and cosmetic sectors.
In addition, DIVERSA goes one step further; our customized nanoemulsions easily allow decoration with proteins and ligands for targeting purposes, taking your research to the next level.
Easy protocols, easy modification, easy research!
We offer ready-to-use kits but also a co-development program. If you need help with your delivery project, we can tailor our technology to your needs and provide testable prototypes within eight weeks!
Our technology is fully compatible with industrial and regulatory scaling, and our co-developments are perfect for in vitro and in vivo validation. We optimize the formulation for the indication of interest!
We want to help you with your project. Contact us to boost your molecule’s delivery!