Producing Lipid Nanoparticles on Do Bio’s Nadia platform

mRNA vaccines such as Pfizer/BioNTech and Moderna were under the spotlight during the COVID-19 pandemic in 2020. The success of these vaccines is remarkable, considering that mRNA, the messenger molecule responsible for giving instructions to the cell to produce a protein, is incredibly delicate and difficult to work with. Delivering mRNA into our cells is tricky due to mRNA being negatively charged, large molecules, which means they cannot simply break through the protective lipid membranes of cells. Nucleic acids are large molecules and require nanoparticle-based technologies or LNPs to shuttle them into cells in the body.

“The Unsung hero”

Lipid nanoparticles (LNPs) have gained traction due to their ability to deliver active pharmaceutical ingredients (APIs) such as nucleic acids (e.g, DNA, mRNA, siRNA…). LNPs come in many different forms including liposomes, solid LNPs, micelles or nanostructure lipid carriers, from which solid LNPs have been developed for use in the Moderna and Pfizer/BioNTech COVID-19 vaccinations. LNP development has been challenging, but without this packaging, mRNA vaccines would not have been possible. “It is the unsung hero of the whole thing,” says Giuseppe Ciaramella, head of infectious diseases at Moderna between 2014 and 2018.

LNPs contain some key ingredients: ionizable lipids whose positive charge binds to negatively charged mRNA, pegylated lipids that help stabilize the nanoparticle, phospholipids and cholesterol molecules that contribute to the structure of the nanoparticle. The concept behind making LNPs involves having these components transport the mRNA cargo, protect it from the body’s immune system and various enzymes, and deliver it to cells. The mRNA is then unloaded which provides the cell with instructions to make the desired protein. The process in theory may seem straightforward, however, in practicality it is challenging.

Why Do LNPs?

The COVID-19 vaccination development and rollout using LNP technology is a good example of why LNPs are important in the world of vaccine therapy. As a result, a variety of lipid nanoparticles have been explored for mRNA delivery. The lessons learnt from clinical studies can be used to further improve mRNA LNP formulations of vaccines, for both existing and emerging pathogens. Delivery of drugs is also an important application of LNPs. Cancer therapy has benefitted from this. The selectivity of LNP encapsulated anti-cancer drugs is superior to that of free drugs. Lipid nanoparticles are believed to be more accessible to some tumors, due to enhanced permeability and retention (EPR) effects, thus, providing higher efficiency in cancer therapy, as it can selectively release anticancer drugs near tumor cells. Much of today’s cancer research focuses on identifying missing or defective genes. LNP-based gene therapy can be used to eliminate the cancer cells and improve the ability of the immune system to recognise and destroy cancer cells, or deliver molecules into cells that can silence inherited mutations.

Devil is in the details

The controlled mixing of the lipids and the mRNA cargo is key in making stable LNPs. In the past, this has been done experimentally using methods like extrusion and solvent injection. These methods are laborious and involve turbulent mixing of fluids. Microfluidic based technologies provide many advantages in comparison. Precise control of parameters like size, and flow rates, control of reaction time, high reproducibility and reduced sample waste are some advantages of producing nanoparticles using microfluidics. Precise control of parameters like size, and flow rates, control of reaction time, high reproducibility and reduced sample waste are some advantages of producing nanoparticles using microfluidics. Besides these benefits, microfluidic platforms such as the Nadia are easy to use and do not require in-depth knowledge of complex microfluidic set-ups.

LNPs with Nadia

Do Bio, a brand of Blacktrace Holdings UK, has leveraged microfluidics for use in the field of Life Sciences. Our sister brand, Dolomite Microfluidics, formed in 2005, has developed microfluidic systems for production of nanoparticles for several years and has an established customer base. More recently, Particle Works has also made its mark within the LNP community with the ALiS and ANP systems. This year, Do Bio will launch its LNP solution for developing biotherapeutics. Nadia users can now, for the first time, reliably produce LNPs on the Nadia platform.

Our instrument range comprising of the Nadia Instrument, Nadia Innovate and Nadia Go, contain three independently controlled pressure pumps which allow different flow rates and flow rate ratios. These can be optimized on the Nadia Innovate and Nadia Go Instruments for adjusting the size of the particle to suit the user’s application needs.

LNPs produced on the Nadia Go and Nadia Innovate using the Nadia chip can then be produced in up to 8 parallel runs on the Nadia Instrument. Temperature control is a unique feature to the Nadia instrument range which can be useful for making LNPs that require an elevated temperature. The technology is agnostic to the chemistry of the formulations. For guidance on how your application needs can fit within our technology, please contact support@dolomite-bio.

All of these features combined means that the Nadia product family has huge potential in the development of LNP based therapeutics and research.

If you’d like to learn more about producing LNPs on the Nadia Innovate, have a look at the Application Note for Lipid Nanoparticle synthesis on the Nadia


LNP synthesis on the Nadia Platform app note


The LNP synthesis on the Nadia Platform intro video