New research by University of Texas at Dallas scientists might help solve a significant problem within the deployment of sure COVID-19 vaccines worldwide — the necessity for the vaccines to be stored at below-freezing temperatures throughout transport and storage.
In a examine revealed on-line April 13 in Nature Communications, the researchers display a brand new, cheap approach that generates crystalline exoskeletons round delicate liposomes and different lipid nanoparticles and stabilizes them at room temperature for an prolonged interval — as much as two months — of their proof-of-concept experiments.
The Moderna and Pfizer/BioNTech COVID-19 vaccines use lipid nanoparticles — principally spheres of fats molecules — to guard and ship the messenger RNA that generates a vaccine recipient’s immune response to the SARS-CoV-2 virus.
“The expense of keeping these vaccines very cold from the time they’re made to the time they’re delivered is a challenge that needs to be addressed, especially because many countries don’t have sufficient infrastructure to maintain this kind of cold chain,” stated Dr. Jeremiah Gassensmith, affiliate professor of chemistry and biochemistry and of bioengineering at UT Dallas and a corresponding writer of the examine. “Although we did not include in this work the specific lipid nanoparticles used in current COVID-19 vaccines, our findings are a step toward stabilizing a lipid nanoparticle in a way that’s never been done before, so far as we know.”
The thought for the research challenge started throughout a coffee-break dialogue between Gassensmith and Dr. Gabriele Meloni, a corresponding co-author of the examine and assistant professor of chemistry and biochemistry within the School of Natural Sciences and Mathematics at UT Dallas.
Gassensmith’s space of experience is biomaterials and metal-organic frameworks, whereas Meloni’s research focus is transmembrane transporter proteins. These proteins reside inside cell membranes and are essential for shifting a wide range of small molecules, together with ions and hint metals, out and in of cells for a number of functions.
“Membrane proteins sit in a cell membrane, which is a lipid bilayer,” Meloni stated. “To study their structure and biophysical and biochemical properties, we must extract these proteins from the membrane using detergents and then reconstitute them back into an artificial membrane — a proteoliposome — that mimics the proteins’ natural environment.”
Lipid nanoparticles and liposomes are comparable in construction, and neither are thermodynamically secure at room temperature, Gassensmith stated. The lipid constructions can fuse or combination, exposing any embedded membrane proteins or cargo to degradation.
“One of the challenges in my field of research is that both membrane proteins and lipid bilayers are very delicate and intrinsically metastable, and we’re trying to combine them in order to understand how these proteins function,” Meloni stated. “We have to handle them carefully and prepare them fresh each time. They cannot be stored for long periods and are not easily shipped to colleagues in other labs.”
The researchers joined forces to develop a strategy to stabilize this type of lipid system and demonstrated their outcomes utilizing transmembrane proteins from Meloni’s lab as a case examine.
They blended liposomes — some with embedded proteins, some with out — with a mix of two cheap chemical compounds, zinc acetate and methylimidazole, in a buffer answer. In a couple of minute, a crystal matrix started to kind round particular person liposomes.
“We think that the lipids interact with the zinc just strongly enough to form an initial zinc-methylimidazole structure that then grows around the lipid sphere and completely envelops it, like an exoskeleton,” Gassensmith stated. “It’s analogous to biomineralization, which is how certain animals form shells. We sort of co-opted nature in creating this totally fake shell, where the biomacromolecules — the lipids and proteins — catalyze the growth of this exoskeleton.”
The capacity of biomimetic shells to kind round organic molecules just isn’t new, Gassensmith stated, however the course of hasn’t labored effectively with lipids or liposomes as a result of the steel salts that comprise the shell materials suck water out of the liposomes by osmosis and trigger them to blow up.
“One of the keys to this research was identifying the buffer solution in which everything resides,” Gassensmith stated.
Building a Buffer
Three graduate college students collaborated on the challenge to develop the distinctive buffer medium that enables the response to happen.
“The buffer medium maintains the ionic strength of the solution and keeps the pH stable so that when you add a huge amount of metal salts, it doesn’t osmotically shock the system,” stated Fabián Castro BS’18, a chemistry doctoral pupil in Gassensmith’s lab and a lead writer of the examine.
Castro and co-lead authors Sameera Abeyrathna and Nisansala Abeyrathna, chemistry doctoral college students (and siblings) in Meloni’s lab, labored collectively to develop the buffer formulation.
Once the biomolecules have grown a shell, they’re locked in, and the lipids stay secure. While the exoskeleton may be very secure, it has a fortuitous Achilles’ heel.
“The shell will dissolve if it encounters something that is attracted to zinc,” Gassensmith stated. “So, to release and reconstitute the liposomes, we used a zinc chelating factor called EDTA (ethylenediaminetetraacetic acid), which is a common, inexpensive food additive and medicine used to treat lead poisoning.”
In addition to the laboratory experiments, in one other proof-of idea train, Gassensmith mailed via the U.S. Postal Service a pattern of the stabilized lipid particles to his mom in Rhode Island. She shipped them again to Texas, however as a result of the COVID-19 pandemic pressured the shutdown of most UT Dallas research labs in 2020, the samples sat untouched for about two months till the graduate college students returned to campus to look at them. Although the casual experiment lasted for much longer than the researchers had anticipated, the samples survived and functioned “just fine,” Gassensmith stated.
“This project required two different types of expertise — my group’s expertise in membrane transport proteins and Dr. Gassensmith’s long track record working with metal-organic frameworks,” Meloni stated. “Our success clearly demonstrates how such collaborative research can bring about novel and useful results.”
Other UT Dallas authors of the examine within the Department of Chemistry and Biochemistry embrace Dr. Ron Smaldone, affiliate professor; doctoral college students Yalini Wijesundara, Olivia Brohlin, Alejandra Durand Silva and Shashini Diwakara; and Michael Luzuriaga PhD’20. Researchers from Graz University of Technology in Austria additionally contributed to the work.
The research was funded partly by the National Science Foundation, National Institute of General Medical Sciences of the National Institutes of Health (R35GM128704), The Welch Foundation, the U.S. Army Combat Capabilities Development Command Army Research Laboratory, the UT Dallas Office of Research’s Seed Program for Interdisciplinary Research, the Mexican National Council of Science and Technology, the European Union’s Horizon 2020 Program, and the Central European Research Infrastructure Consortium.