|MIT Department: Biological Engineering
Faculty Mentor: Prof. Darrell Irvine
Undergraduate Institution: University of Florida
Originally from Caracas, Venezuela, I am pursuing a degree in biomedical engineering at the University of Florida, with an interest in computer science. My motivation for this comes from trying to understand how we can mobilize and use the field of software technology to aid in lessening healthcare disparities worldwide, and plan on pursuing a joint MD/PhD program. As part of the Rinaldi-Ramos Lab at UF, I have worked on and learned about the applications of magnetic nanoparticles designed for magnetic particle imaging as both a diagnostic and therapeutic tool in biomedical applications. My hobbies include swimming and triathlon training, with the hopes of one day completing a full Ironman.
Cryoprotectant and Storage Conditions for Lipid Nanoparticle Formulations
Manuel Cortes1, Byungji Kim2, Ryan Hosn2, and Darrell J. Irvine2,3,4
1Department of Biomedical Engineering, University of Florida
2Department of Biological Engineering, Massachusetts Institute of Technology
3Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
4Howard Hughes Medical Institute
The ability to maintain vaccine efficacy from synthesis, to storage, and lastly administration is among the most important clinical-setting considerations. Due to the enzymatic susceptibility of RNA vaccine technology, optimization of the cryoprotectant and storage conditions is an essential step. Here, we explore sucrose as a cryoprotectant with varying concentrations (0%, 5%, 10%, 30% in saline), solvents (phosphate-buffered saline and tris-buffered saline), and storage temperatures (4°C, -20°C, -80°C, and liquid nitrogen) for an RNA-loaded lipid nanoparticle (LNP) formulation for HIV. In vitro data from dynamic light scattering (DLS) and cryogenic electron microscopy (Cryo-EM) narrowed down the leading storage temperatures to 4°C and -20°C, as they maintained a polydispersity and hydrodynamic diameter comparable to the freshly synthesized batch. For the in vivo experiments, IVIS bioluminescent imaging showed PBS saline to provide the strongest signal, while LNPs with a concentration of 10% sucrose exhibited the strongest immunological activity using an enzyme linked immunosorbent assay (ELISA). Consolidating these results, we proved that storage at -20°C in PBS at a 10% sucrose concentration optimizes RNA-LNP stability and immunological activity. Overall, this study has important implications in achieving sustainable production and distribution goals for vaccines in a global setting.