HDX Solution

HDX Solution – unlock insights in protein structure conformation

  • Are you looking for a way to identify structural hotspots within your potential biotherapeutic protein?

  • Are you searching for a solution for understanding structural conformational changes of your biotherapeutic during formulation or storage?

  • Do you want to understand more about the protein-ligand (drug/protein) interactions and their relationship to drug activity?

The Bruker HDX Solution enables you to answer these questions and unlock insights into protein structure conformation, delivering reliable end results - first time, every time.

Secondary Structure Representation of the crystal structure of E.coli HtpG colour coded according to H/D exchange at 30s
Secondary Structure Representation of the crystal structure of E.coli HtpG colour coded according to H/D exchange at 30s (click to enlarge)

The Challenges of Biotherapeutic Development

Researchers within (bio)pharmaceutical sciences are increasingly tasked with gaining a deeper knowledge and understanding of the effects interactions and stressors such as pH or temperature, have on the conformational structure of biotherapeutic proteins. This is a very challenging task and researchers often use a variety of analytical tools to study the higher order structure of the biotherapeutics, monitor protein/protein-drug interactions and perform epitope mapping. 

In recent years Hydrogen Deuterium Exchange Mass Spectrometry (HDX-MS) has emerged as a valuable technique within the arsenal of analytical tools available for providing such insights into protein structure conformation. However, until recently HDX-MS was confined mostly to academic research laboratories, due to a lack of automation and limited data interpretation tools, as well as the need to have reliable monoisotopic mass assignments.

Benefits of Combining Automated HDX with UHR-QTOF MS Technology

Key components of the robotic setup for automatic HX-MS experiments
Key components of the robotic setup for automatic HX-MS experiments (click to enlarge)

Due to the high level of specificity and sensitivity achieved in both MS and MS/MS the use of Ultra-High Resolution Quadrupole Time-of-Flight (UHR-QTOF) technology for biotherapeutic charaterization is well-established. The ability of UHR-QTOFs to provide correct monoisotopic accurate-mass readout whilst maintaining true isotopic patterns is necessary for achieving accurate, reliable H/D ratios enabling Researchers to fully explore the quantitative results from HDX-MS studies.

The ability to couple accurate monoisotopic peptide mass information with data obtained from innovative fragmentation approaches such as Electron Transfer Dissociation (ETD) on the UHR-QTOF platform, provides additional insights. No longer do we just know which peptide regions are affected by the perturbation but we can now pinpoint the changes occurring at the residue level.  

However, for HDX-MS to be accepted as a routine, reliable technology within the (bio)pharmaceutical sciences, the ability to readily interpret the data obtained and automate the entire HDX-MS time-course experiment from sample preparation through to LC-MS analysis is necessary.

HDX Solution Providing Fast, Automated, Reliable, Quantitative Structural Insights

The Bruker HDX Solution combines the LEAP HDx PAL, with the market-leading Bruker maXis II ETD and state of the art software from Sierra Analytics (HDExaminer) for data analysis. The industry-leading ETD capabilities of Bruker’s maXis II ETD UHR-QTOF offers unparalleled sequence coverage for middle-down and top-down studies while maintaining access to your existing peptide mapping workflows. However, the evolution of HDX-MS from bottom-up strategies based upon pepsin-digestion toward middle-down and top-down studies utilizing the power of ETD fragmentation has the potential to revolutionize the way conformational studies are performed.

The Bruker HDX Solution tool kit delivers automated and reliable HDX-MS workflows with correct monoisotopic accurate-mass readout to fully explore quantitative results from H/D ratios. This enables structural insights about biomolecules to be accessed reliably and with greater certainty and confidence. 

Click here to see the Technical Details or to see the latest Applications.

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