Epitope Mapping

Dr. Anne Zeck
Group Leader Bioanalytics

Verification and determination of the exact binding region of antibodies by HDX-LC-MS

We offer characterization of conformational epitopes by hydrogen-deuterium exchange with mass spectrometry analysis (HDX-MS). By complexing the antibody with the antigen and then deuteration of the solvent-accessible amide protons, the increase in weight of the antibody-bound and free antigens allows for epitope regions to be mapped. To further increase the resolution of this method, the molecules are hydrolysed with pepsin to form small peptides prior to analysis, which can then be chromatographically separated at low temperatures and acidic pH conditions and then detected in the mass spectrometer.


Our services

  • Epitope mapping of conformational epitopes by HDX-LC-MS



  • Low temperature LC
  • MS
  • Peptide Mapping
  • ES-MS
  • ES-MS/MS



  • Gramlich M, Hays HCW, Crichton S, Kaiser PD, Heine A, Schneiderhan-Marra N, Rothbauer U, Stoll D, Maier S, Zeck A. HDX-MS for Epitope Characterization of a Therapeutic ANTIBODY Candidate on the Calcium-Binding Protein Annexin-A1. Antibodies (Basel). 2021 Mar 19;10(1):11. doi: 10.3390/antib10010011. PMID: 33808657; PMCID: PMC8006148.
  • Wagner TR, Ostertag E, Kaiser PD, Gramlich M, Ruetalo N, Junker D, Haering J, Traenkle B, Becker M, Dulovic A, Schweizer H, Nueske S, Scholz A, Zeck A, Schenke-Layland K, Nelde A, Strengert M, Walz JS, Zocher G, Stehle T, Schindler M, Schneiderhan-Marra N, Rothbauer U. NeutrobodyPlex-monitoring SARS-CoV-2 neutralizing immune responses using nanobodies. EMBO Rep. 2021 May 5;22(5):e52325. doi: 10.15252/embr.202052325. Epub 2021 Apr 27. PMID: 33904225; PMCID: PMC8097376.