Epitope mapping is performed to identify the epitope to which a monoclonal antibody binds.
Conformational epitopes consist of residues that are scattered in the primary sequence but brought together through folding of the protein of interest. The determination of conformational epitopes is a major issue in the antibody field, the standard techniques for determining linear epitopes being ineffective.
Agro-Bio has therefore developed a robust method for identifying an antibody’s conformational epitopes using in situ proteolysis involving formation of an immobilized antigen-antibody complex, followed by mass spectrometry analysis.
Our method combines:
Two approaches are possible:
Identification of the sequence of interest is based on the fact that antigen/antibody complex formation protects the epitope from enzyme digestion.
After affinity chromatography and digestion, the epitope fragments are eluted then sequenced by mass spectrometry: MALDI-TOF/TOF.