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The SEC method can also be used to measure binding constants, effects of cofactors on binding, competitive and noncompetitive binding, and other ligand interactions with target proteins.
Affinity Competition Experiments enable protein-ligand binding affinity measurements in compound mixtures. As simulated in (A), a library of compounds of varying affinity (blue) is embedded with calibrant ligands of known Kd (red) and titrated with a compound of known Kd (purple) to yield MS-measured ACE50 curves. (B) A calibration curve generated from the calibrants' ACE50 and Kd values yields the other mixture components Kds.
from: D.A. Annis et al., Anal. Chem. 2007, 79, 4538-4542.