Conformational Assessment of Adnectin and Adnectin-Drug Conjugate by Hydrogen/Deuterium Exchange Mass Spectrometry.
Richard Y-C HuangSteven R O'NeilDaša LipovšekGuodong ChenPublished in: Journal of the American Society for Mass Spectrometry (2018)
Higher-order structure (HOS) characterization of therapeutic protein-drug conjugates for comprehensive assessment of conjugation-induced protein conformational changes is an important consideration in the biopharmaceutical industry to ensure proper behavior of protein therapeutics. In this study, conformational dynamics of a small therapeutic protein, adnectin 1, together with its drug conjugate were characterized by hydrogen/deuterium exchange mass spectrometry (HDX-MS) with different spatial resolutions. Top-down HDX allows detailed assessment of the residue-level deuterium content in the payload conjugation region. HDX-MS dataset revealed the ability of peptide-based payload/linker to retain deuterium in HDX experiments. Combined results from intact, top-down, and bottom-up HDX indicated no significant conformational changes of adnectin 1 upon payload conjugation. Graphical Abstract ᅟ.
Keyphrases
- mass spectrometry
- molecular dynamics
- molecular dynamics simulations
- single molecule
- protein protein
- amino acid
- liquid chromatography
- multiple sclerosis
- binding protein
- high resolution
- ms ms
- cancer therapy
- gas chromatography
- drug induced
- high performance liquid chromatography
- adverse drug
- emergency department
- high glucose
- endothelial cells
- solid phase extraction