Probing Antibody Structures by Hydrogen/Deuterium Exchange Mass Spectrometry.
Zuzana KalaninováLukáš FojtíkJosef ChmelíkPetr NovákMichael VolnyPetr ManPublished in: Methods in molecular biology (Clifton, N.J.) (2023)
Hydrogen/deuterium exchange (HDX) followed by mass spectrometry detection (MS) provides a fast, reliable, and detailed solution for the assessment of a protein structure. It has been widely recognized as an indispensable tool and already approved by several regulatory agencies as a structural technique for the validation of protein biopharmaceuticals, including antibody-based drugs. Antibodies are of a key importance in life and medical sciences but considered to be challenging analytical targets because of their compact structure stabilized by disulfide bonds and due to the presence of glycosylation. Despite these difficulties, there are already numerous excellent studies describing MS-based antibody structure characterization. In this chapter, we describe a universal HDX-MS workflow. Deeper attention is paid to sample handling, optimization procedures, and feasibility stages, as these elements of the HDX experiment are crucial for obtaining reliable detailed and spatially well-resolved information.
Keyphrases
- mass spectrometry
- liquid chromatography
- high resolution
- gas chromatography
- high performance liquid chromatography
- capillary electrophoresis
- multiple sclerosis
- ms ms
- tandem mass spectrometry
- transcription factor
- healthcare
- amino acid
- social media
- loop mediated isothermal amplification
- simultaneous determination
- visible light
- drug administration