Messenger RNA (mRNA) sequences can be
engineered to encode certain proteins that can be translated and expressed
within cells. Lipid nanoparticles (LNPs) are used to package mRNA payload for
delivery in vaccines. The four lipid components
of LNPs are an ionizable cationic lipid, a neutral helper lipid, cholesterol,
and a PEGylated lipid. My research focuses on the synthesis and
purification of ionizable lipids used to formulate LNPs. The storage temperature and time stored play a large
role in the stability of LNPs as the mRNA can degrade, rendering the LNPs
ineffective. Research suggests that the lipid components of the LNPs may
degrade over time as well1. Over the past semester, I have
developed a method to test the stability of the lipid components of LNPs stored
at 4°C, 25°C, and 37°C.
After LNPs are formulated, the encapsulated mRNA can be extracted
from the nanoparticles for analysis. The extraction method results in an
aqueous phase containing the mRNA, as well as an organic phase containing the
four lipid components. For my project, I examined the extracts from LNPs stored
at 4°C, room temperature (25°C), and 37°C over
the course of 3 weeks.
For each week of the 3-week stability study, I
used thin layer chromatography (TLC) to qualitatively examine the aqueous and
organic extracts from the LNPs stored at each temperature. Liquid
chromatography mass spectrometry (LCMS) spectra were analyzed to observe the
fragmentation of the compounds. Nuclear magnetic
resonance (NMR) spectra were also used to examine the molecular structure of
the compounds.
A typical day of research is arriving at the lab and checking on synthesis reactions set up the night before. Typically I will run three columns each day as well as run my samples on the LCMS. My research experience has been exciting, challenging, and rewarding. I am interested in the field of bioengineering so this project fits into my future plans of attending graduate school.
1.)European Medicines Agency, Committee for Medicinal Products for Human Use (CHMP); Assessment Report, Covid-19 Vaccine Moderna. 2021
