Fig. 3
From: A novel in-vitro expression assay by LC/MS/MS enables multi-antigen mRNA vaccine characterization
mRNA Vaccine Dose Response. The mRNA vaccine dose response, encapsulating the efficiency of mRNA transfection into HEK293T cells and subsequent expression of the encoded antigen proteins, was measured by IVE-LC/MS/MS and IVE-flow cytometry. HEK293T cells were dosed with a quadrivalent mRNA vaccine containing an equal theoretical load of each mRNA construct. The resulting data from the IVE-LC/MS/MS assay are graphed as % antigen peptide signal for each unique antigen relative to specific HEK293T peptide signal vs. total mRNA dosed (ng) per plate well (R2 for Wisconsin, Darwin, Austria and Phuket are: 0.963, 0.985, 0.99 and 0.980 respectively) (A). A linear regression of these same data in pmol antigen per pellet is plotted in B (R2 for Wisconsin, Darwin, Austria and Phuket are: 0.992, 0.970, 0.997 and 0.995 respectively). IVE-flow cytometry assay results are graphed as % cells positively expressing the antigen protein of interest (C) and mean fluorescence intensity (MFI) ratio (D) vs. the total mRNA (ng) per plate well.
