Probing Antibody Structures by Hydrogen/Deuterium Exchange Mass Spectrometry

Kalaninová, Zuzana; Fojtík, Lukáš; Chmelík, Josef; Novák, Petr; Volný, Michael; Man, Petr

doi:10.1007/978-1-0716-3457-8_17

Cited by 2 publications

(1 citation statement)

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“…To improve the sequence coverage of HER2, we utilized online deglycosylation and used quench conditions compatible with PNGase Rc . They are however milder than required for proper denaturation and reduction of antibodies, and thus, the antibody coverage is compromised, especially in the N-teminal half of the light chain and part of the heavy chain . Therefore, the antibody deuteration profile was not followed, and the conditions where the antibody is alone were not included.…”

Section: Resultsmentioning

confidence: 99%

Structural Characterization of Monoclonal Antibodies and Epitope Mapping by FFAP Footprinting

Fojtík,

Kalaninová,

Fiala

et al. 2024

Anal. Chem.

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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.

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Section: Resultsmentioning

confidence: 99%