Scientists in Germany develop a new analytical method, which enables improved insight into (mRNA) nanoparticles and similar pharmaceutical and non-pharmaceutical products

Messenger RNA (mRNA) nanomedicines, a ground-breaking technology that has led to the development of the first approved COVID-19 vaccine, was recently recognised by the Nobel Prize in Medicine or Physiology. But mRNA’s potential for pharmaceutical application is expected to go much beyond this – it could open up new opportunities for the treatment and prevention of diseases, such as viral and bacterial infections, cancer, cardiovascular diseases, and inflammatory and auto-immune diseases. It could also transform the large field of interventions by therapeutic proteins.

Many novel mRNA nanomedicines, which are currently in different stages of development, may become available in the future. One requirement for all applications of mRNA in pharmaceutical products is that they need to be formulated in suitable delivery systems, each designed for different functions and optimised for therapeutic product needs based on the intended application and route of delivery.

Lipid-based nanoparticles are tiny droplets of fat-like molecules that serve as protective packaging for the mRNA. Their properties depend on composition, structure, manufacturing protocol, and other conditions. An important aspect of nanoparticles is their size. By their nature, nanoparticles can vary a little bit in size, some being a bit smaller, some a bit larger than the average value. The particle size can have an influence, for example, on the stability and the behaviour of the formulations after administration. It is therefore important to control the particle size inside a pharmaceutical product to evaluate and ensure its quality.

Scientists at EMBL Hamburg, Johannes Gutenberg University Mainz, Postnova Analytics GmbH, and BioNTech SE have developed a new method to precisely elucidate the size of all particles in such pharmaceutical products, as well as their structure and how many RNA molecules they carry inside them. The study was conducted based on lipoplex formulations, a mRNA delivering technology developed by BioNTech.

“So far, it was very difficult to measure all these size-related properties; therefore, often only average values were determined,” said Heinrich Haas, one of the leaders of the project. “With our new method, we can determine many size-related features all at once, with a single measurement and for all nanoparticles in a product. This information can be very useful to evaluate product quality.”

The method will also be applicable for the investigation of other pharmaceutical products.

“Liposomes are another type of pharmaceutical nanoparticles which have been applied since years for treatment of cancer or infectious diseases such as fungal infections,” said Peter Langguth, the project leader at Johannes Gutenberg University Mainz. “Now even generic liposome products are available on the market, and probably there will be more to come. The new method can be very useful in evaluating the quality of these generics in comparison to the originator products and will pave the way for further high-quality drug products at an even more reasonable cost.”

A two-in-one method

What makes the new method so powerful is that it couples two techniques: asymmetrical-flow field-flow fractionation (AF4) and small-angle X-ray scattering (SAXS). AF4 separates lipid-based nanoparticles from other parts of an mRNA nanomedicine and sorts them according to their size. SAXS allows scientists to determine the structure and the number of the sorted particles. To do this unequivocally, it is necessary that only one type of particles is analysed at a time, which is why combining sorting and measuring is so critical.

SAXS is one of the key techniques applied and available at EMBL Hamburg as a service for researchers from academia and industry in Europe and beyond. EMBL Hamburg’s SAXS beamline at the PETRA III synchrotron, now equipped with the AF4 device – set-up with the help of collaborators at Postnova Analytics GmbH – will open up new opportunities not only for studying pharmaceutical nanoparticles, but also for other types of research.

"The combination of these two tools can now be used in many different areas of science,” said Melissa Graewert, Staff Scientist at EMBL Hamburg. “In addition to helping create new medicines, we can also use them to understand how different-sized particles interact in complex biological systems. For example, I've now used this new setup to closely examine how very small plastic debris called nanoplastics, which pollute our waters, can be covered by binding proteins on their surface. A key question is whether this protein shielding enables nanoplastics to travel through our bloodstream, potentially reaching different organs, as they may no longer be recognised as foreign objects by our immune system.”

This work follows up on several previous collaborative studies between EMBL Hamburg, BioNTech SE, and Johannes Gutenberg University Mainz, which explored how mRNA can be better formulated and delivered into human cells. The scientists are continuing their collaborative research to further explore the application of mRNA nanomedicines.

Funding

This research was funded by the "Bundesministerium für Bildung und Forschung BMBF“ grant 05K22UM3 and by the “Deutsche Forschungsgemeinschaft DFG” as part of the collaborative research center (CRC) 1066.

About EMBL

The European Molecular Biology Laboratory (EMBL) is Europe’s life sciences laboratory. We provide leadership and coordination for the life sciences across Europe, and our world-class fundamental research seeks collaborative and interdisciplinary solutions for some of society’s biggest challenges. We provide training for students and scientists, drive the development of new technology and methods in the life sciences, and offer state-of-the-art research infrastructure for a wide range of experimental and data services.

EMBL is an intergovernmental organisation with 28 member states, one associate member, and two prospective members. At our six sites in Barcelona, Grenoble, Hamburg, Heidelberg, Hinxton near Cambridge, and Rome, we seek to better understand life in its natural context, from molecules to ecosystems.

About Johannes Gutenberg University Mainz

Johannes Gutenberg University Mainz (JGU) is a globally recognized research-driven university with around 30,000 students from over 120 nations. Its core research areas are in particle and hadron physics, the materials sciences, and translational medicine. JGU's success in Germany's Excellence Strategy program has confirmed its academic excellence: In 2018, the research network PRISMA+ (Precision Physics, Fundamental Interactions and Structure of Matter) was recognized as a Cluster of Excellence – building on its forerunner, PRISMA. Moreover, excellent placings in national and international rankings as well as numerous honors and awards demonstrate the research and teaching quality of Mainz-based researchers and academics. Further information at www.uni-mainz.de/eng

About BioNTech

Biopharmaceutical New Technologies (BioNTech) is a next generation immunotherapy company pioneering novel therapies for cancer and other serious diseases. The Company exploits a wide array of computational discovery and therapeutic drug platforms for the rapid development of novel biopharmaceuticals. Its broad portfolio of oncology product candidates includes individualized and off-the-shelf mRNA-based therapies, innovative chimeric antigen receptor (“CAR”) T cells, several protein-based therapeutics, including bispecific immune checkpoint modulators, targeted cancer antibodies and antibody-drug conjugate (“ADC”) therapeutics, as well as small molecules. Based on its deep expertise in mRNA vaccine development and in-house manufacturing capabilities, BioNTech and its collaborators are developing multiple mRNA vaccine candidates for a range of infectious diseases alongside its diverse oncology pipeline. BioNTech has established a broad set of relationships with multiple global pharmaceutical collaborators, including Duality Biologics, Fosun Pharma, Genentech, a member of the Roche Group, Genevant, Genmab, OncoC4, Regeneron, Sanofi and Pfizer.

For more information, please visit www.BioNTech.com.

About Postnova Analytics

Postnova is the inventor and leader in field-flow fractionation and an innovator in light scattering technology, offering instruments, software, and services used in laboratories of universities, institutions, and corporations worldwide to separate and characterize analytes in the nanometer to micrometer and kilodalton to gigadalton range. In a constantly growing number of applications around polymers and further advanced materials, nutrition, and personal care, as well as in environmental and pharmaceutical science, Postnova's offering and expertise help its customers to develop and optimize products and their production processes, to ensure product quality, to contribute to a safer and more sustainable environment, as well as to fight and cure diseases.

Information about Postnova is available at www.postnova.com.