Structural Characterization of Monoclonal Antibodies and Epitope Mapping by FFAP Footprinting.
Lukáš FojtíkZuzana KalaninováJan FialaPetr HaladaJosef ChmelíkPetr ManZdeněk KukačkaPetr NovákPublished in: Analytical chemistry (2024)
Covalent labeling in combination with mass spectrometry is a powerful approach used in structural biology to study protein structures, interactions, and dynamics. Recently, the toolbox of covalent labeling techniques has been expanded with fast fluoroalkylation of proteins (FFAP). FFAP is a novel radical labeling method that utilizes fluoroalkyl radicals generated from hypervalent Togni reagents for targeting aromatic residues. This report further demonstrates the benefits of FFAP as a new method for structural characterization of therapeutic antibodies and interaction interfaces of antigen-antibody complexes. The results obtained from human trastuzumab and its complex with human epidermal growth factor receptor 2 (HER2) correlate well with previously published structural data and demonstrate the potential of FFAP in structural biology.
Keyphrases
- epidermal growth factor receptor
- endothelial cells
- mass spectrometry
- high resolution
- tyrosine kinase
- advanced non small cell lung cancer
- induced pluripotent stem cells
- pluripotent stem cells
- amino acid
- systematic review
- electronic health record
- machine learning
- small molecule
- ms ms
- high performance liquid chromatography
- monoclonal antibody
- simultaneous determination