In partnership with Trajan, Waters has transformed a highly manual analytical process into an integrated and automated system with our HDX-MS system, which leverages high-pressure nano- to micro-scale UPLC separations, sample handling robotics, and high-resolution, ion mobility mass spectrometry for in-depth analysis of protein structure studies, including:
Waters commercially-available complete system for HDX-MS studies pairs our ACQUITY UPLC M-Class System with HDX Technology with a Waters mass spectrometer like the SELECT SERIES Cyclic IMS, Trajan robotics and HDX application-specific software, DynamX.
Hydrogen deuterium exchange (HDX) MS works in many applications, including understanding how small-molecule therapeutics bind to protein targets, and performing epitope mapping. With this technology, deuterium in a solution replaces the amide hydrogen in a biomolecule’s backbone, and MS measures these changes.
Amide hydrogen on the backbone of a biomolecule exchanges with deuterium in solution at different rates partly dependent on conformation. MS can measure these changes and therefore infer the degree of activity at various sites. These locations can be identified and mapped to the primary sequence, which allows for easier visualization and reveals information about protein structure. Relative folding and dynamics can be determined from the different rates of uptake at different locations.
With UPLC-MSE, your lab can generate a comprehensive dataset, measuring all peptides without bias by peptide signal intensity. This is an important feature for HDX studies, where key information might be found in low-intensity peptides.
HDX-MS opens new windows into biomolecule dynamics by providing information on the relative deuterium uptake of different conformations of a protein, or locations within a protein. Recent developments have made HDX with mass spectrometry a more accessible tool to study the dynamics of higher order structure (HOS) of biotherapeutics. Industry and regulatory trends have clearly moved towards an emphasis on conformation and the relationships between biomolecules, making HDX a key tool being rapidly adopted in more biotherapeutic analysis areas.
Biosimilar Characterization
Leverage UPLC separations and high-resolution mass spectrometry (HRMS) to confidently identify changes in protein conformation with the ACQUITY UPLC M-Class System with HDX Technology.
Maximize sample information for your analytes from detailed characterization to accurate quantitation with the Xevo G3 QTof, which enables complete confidence in your results.
Developed through focused collaboration, the SELECT SERIES Cyclic IMS combines novel cyclic ion mobility separation with state-of-the-art high performance, time-of-flight mass spectrometry, enabling leading researchers to unlock the potential in scientific discovery.
Achieve ultimate flexibility and greater freedom of analytical choice to support scientific creativity and technical success for any application with SYNAPT XS.
Deuterium retention for four peptides from yeast enolase tryptic peptides.
Crystal structure of CaM color coded to represent deuterium uptake. (A) Apo CaM and (B) holo CaM at 10 s deuterium labeling were compared. A significant difference in deuterium uptake was observed in the highlighted region where the conformation has been changed due to the calcium binding.
Different deuterium uptake of the same peptide was found in Apo and Holo CaM. At 10-minute labeling, this peptide (FKEASLF, 12-19) showed a higher deuterium uptake in Apo (left panel) than in Holo (right panel).
Comparison of deuterium loss from fully deuterated bradykinin and angiotensin II at A) various digestion temperatures, B) quench holding times, and C) digestion/desalting flow rate. The deuterium loss caused by mass spectrometer source is highlighted with a black border in 5B. Similar isotope profiles were found in both peptides under normal pressure and high pressure (5D). High pressure caused a slightly higher deuterium loss (<5%).
