Off-target quantification

Valid ated off-target candidates can be quantified on cells and tissues by mass spectrometry. Heavy-labelled reference peptides are obtained and spiked at a known concentration into HLA peptide eluates from cells and tissues before mass spectrometric analysis. Comparing the signal intensity of the endogenous and the reference peptide through highly sensitive PRM mass spectrometric assays facilitates the calculation of off-target peptide density on cells. With the measured affinity and the density of the HLA peptide complexes, an evaluation of the safety risk can be performed and regulatory authorities can be convinced about the safety of the experimental drug candidate.

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