Hydrogen/deuterium exchange mass spectrometry applied to IL-23 interaction characteristics: potential impact for therapeutics

Abstract

Interleukin-23 (IL-23) is an important therapeutic target for the treatment of inflammatory diseases. Adnectins are targeted protein therapeutics that are derived from domain III of human fibronectin, and have similar protein scaffold to antibodies. A specific adnectin (Adnectin 2) was identified to bind to IL-23 and compete with IL-23/IL-23R interaction, being a potential protein therapeutic. Hydrogen/deuterium exchange mass spectrometry (HDX MS) and computational methods were applied to probe the binding interactions between IL-23 and Adnectin2 and to determine the correlation between the two orthogonal methods. This review article summarizes the current structural knowledge about Il-23 and it focuses on the applicability of HDX MS to investigate the higher order structure of proteins, which plays an important role for the discovery of new and improved biotherapeutics.

Cutting-edge multi-level analytical and structural characterization of antibody-drug conjugates: present and future

“…Several techniques have been conducted to provide more insights about higher order structures (HOS) of mAbs and ADCs, among which hydrogen-deuterium exchange followed by MS analysis (HDX-MS) is particularly used [163]. Differences in isotope exchange kinetics of amide hydrogens in mAbs have been successfully analyzed to assess the structural impact of post-translational modifications (PTMs) [164,165] while other groups have used this technique to characterize the mAb/Ag interaction (epitope mapping) [166][167][168]. HOS of mAbs might also be affected by the addition of cytotoxic molecules that could potentially give rise to some undesired effects in terms of safety and efficiency.…”

HD-eXplosion: visualization of hydrogen–deuterium exchange data as chiclet and volcano plots with statistical filtering

“…Data interpretation however is complicated, as uneven loss of deuterium and difficulty in creating a fully deuterated control, limit the ability to measure underlying rates of exchange. HDX is more often used comparatively, looking at a protein in multiple states, such as with and without a ligand (Chalmers et al, 2006(Chalmers et al, , 2011Iacob et al, 2015). In these experiments, the data must be interrogated to determine if deuterium uptake of a peptide varies between states (Hageman and Weis, 2019).…”

Content Is King: Databases Preserve the Collective Information of Science

“…Cryo-electron microscopy is now the premier method for this task, but MS-centered strategies, such as crosslinking, hydrogen/deuterium amide exchange, fast photochemical oxidation of proteins, and stability of proteins from rates of oxidation, are increasingly used to assess the folding of proteins. [31][32][33][34][35] Whereas these methods currently do not have the resolution of X-ray crystallography or cryoelectron microscopy, the capability of these methods will grow with time, and the information generated should be captured in databases and integrated with all other information about sequences and modifications.…”