RNA therapeutics – tips for production, LNPs, liposomes and detection
Covid-19 vaccines, Nobel prize in medicine 2023, what comes next?
Current clinical trials for mRNA vaccines are targeting melanoma, respiratory syncytial virus (RSV), seasonal influenza and genital herpes. BioNordika can cover researchers puzzling with pre-clinical stage of mRNA vaccine research also in GMP gradient.
BioNordika’s method tips for different stages of mRNA vaccine development:
1. How to synthetize suitable RNA?
Important things to consider when producing mRNA for therapeutics and vaccines is capping and if the mRNA can eventually easily be synthesized in GMP-grade. Cap-0 is required for efficient translation of the RNA in eukaryotic systems. Our partner’s New England Biolabs (NEB) has developed several different RNA synthesis kits that can be bought over the counter or GMP-grade. Capping alternatives are CleanCap and Vaccinia. Vaccinia is available as GMP-grade reagent if needed.
Some product tips for getting started with RNA synthesis:
HiScribe T7 mRNA Kit with CleanCap® Reagent AG (includes capping) (#E2080)
HiScribe™ T7 ARCA mRNA Synthesis Kit (includes capping and tailing) (#E2060)
HiScribe T7 ARCA mRNA Synthesis Kit (includes capping) (#E2065)
HiScribe™ T7 High Yield RNA Synthesis Kit (#E2040)
HiScribe T7 Quick High Yield RNA Synthesis Kit (#2050)
HiScribe™ SP6 RNA Synthesis Kit (#E2070)
Vaccinia Capping System (#M2080S)
2. Delivery
RNA therapeutics can be transferred in different ways. Most popular ways are to pack RNA molecules to lipid nanoparticles (LNPs) or in lipoplexes which protect the sensitive RNA molecules and help with getting them to the right location.
Developing a correct LNP formulation for your molecules
Packing – lipid nanoparticles (Cayman Chemicals)
Different lipids work better in packing different biomolecules. Ionizable cationic lipids, helper lipids, sterol lipids and PEGylated lipids in different. To learn more about how to work with lipids and what to consider, contact henna.salo@bionordika.se for a PDF copy of LNP guides.
Mouse experiments: In vivo jetRNA+ (PolyPlus)
In the early stages of mRNA therapeutics development, a fast proof of concept alternative for animal studies is in vivo jetRNA+ from PolyPlus. It is liposome based ready-to-use reagent, which is simple to use and can be administered to mice intravenous, intramuscular, intraperitoneal, and retro-orbital injections.
3. Detection of uptake
ModDetect™ Panels (Rockland)
The classic way to detect oligonucleotides in cells is ISH-type assays, but this is often difficult and time consuming with short oligo sequences. One of the common modifications of therapeutic RNA and DNA is phosphorothioate replacement of the natural phosphodiester bond, which Rockland has used as a target in developing it’s immune assay based variant for oligonucleotide therapeutics detection.
More modification detection panels are to come!
The benefit of using the ModDetectTM Panels is the cost-effectiveness. In screening of large oligonucleotide libraries can save 9-12 months in the drug development process in comparison to setting up ISH detection for individual oligos in the library.
Are you working with RNA therapeutics or other nucleic acid therapeutics?
We have a wide selection of products covering research use only and GMP needs. Get in touch with our product specialists through info@bionordika.se to discuss suitable products for your needs.
Henna Salo, PhD
Product Specialist
Stockholm, Uppsala and North Sweden
