Development of a General Purpose Targeted LC-MS Method for Accurate Quantification of the SARS-CoV-2 Spike Protein Expression
Almey, R.; Mwangi, K.; De Cae, S.; Ramasamy, P.; Van Hulle, M.; Lentacker, I.; Schepens, B.; Deforce, D.; Martens, G.; Ramaut, P.; Vissers, J. P.; Saelens, X.; Verbeke, R.; Dhaenens, M.; Van Puyvelde, B. R.
Show abstract
The COVID-19 pandemic has catalyzed interest in immuno-multiple reaction monitoring (immuno-MRM) methods, with the detection of peptides unique to the nucleocapsid protein in nasopharyngeal swabs. While current applications predominantly focus on disease biomarkers, the pandemic has unveiled new opportunities, namely for the quantification of antigen expression following mRNA vaccination. Here, we present an optimized immuno-MRM method for quantifying SARS-CoV-2 spike protein fusion peptide, SFIEDLLFNK, for several practical applications. The method is versatile, applicable to multiple biological matrices, including plasma, and can be extended to nasopharyngeal swabs. It also offers a high-precision tool for assessing protein expression following plasmid and mRNA transfection. Moreover, in parallel to enabling accurate antigen quantification, the flow-through can be used to determine the proteome profile of the infected cells, providing insights into the intracellular immune response. This dual capability supports the rapid optimization of mRNA vaccines, thereby driving advancements in vaccine development strategies.
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