HDX Solution

The Bruker HDX Solution for Automated Structural Conformation Characterization

The Bruker HDX Solution is a complete toolkit combining the LEAP HDx PAL, UHPLC pump, the Bruker maXis II ETD and Sierra Analytics HDExaminer software. 

By utilizing the most advanced (MS) vendor independent HDX system, the LEAP HDx PAL, robustness and reliability are built-in as standard. The LEAP HDx PAL automates the HDX experimental workflow, schedules and performs labelled experiments, followed by quenching and injection under cold conditions.

Innovative, Intelligent Sample Scheduling

One of the key features in bringing HDX-MS into the age of automation and into (bio)pharmaceutical laboratories is the innovative scheduling software, Chronos, included with the LEAP HDx PAL.

 

This software maximizes the chromatographic throughput and hence lab productivity by the intelligent organization and scheduling of the sample preparation steps including labelling, incubation, digestion and quenching.

Optimized Scheduling afforded by the Chronos software provided with the LEAP HDx PAL
Optimized Scheduling afforded by the Chronos software provided with the LEAP HDx PAL (click to enlarge)

High Quality Chromatographic and MS Data

Representative Heat Map produced in Sierra Analytics HDExaminer software showing the differences in deuterium uptake after incubation for 5 s to 86400s
Representative Heat Map produced in Sierra Analytics HDExaminer software showing the differences in deuterium uptake after incubation for 5 s to 86400s (click to enlarge)

Chromatographic separation under UHPLC conditions followed by mass spectrometric detection using the Bruker maXis II with its sub-ppm mass accuracy, high sensitivity and isotopic fidelity guarantees, high quality data and ultimately confidence in the H/D ratios.

 

Market-leading ETD capabilities open-up a new era in HDX-MS experiments. The ability to enhance knowledge by probing at the residue level not just the peptide level potentially gives additional specificity and highlights potential hotspots more readily. Top-down and middle-down approaches are also possible and provide complementary data to that obtained from traditional bottom-up pepsin-digestion based HDX-MS experiments.

 

The Most-widely Used HDX-MS Data Interpretation Software


The ability to interpret and analyze the mass spectral data providing a comprehensive picture of the deuteration uptake behaviour of a protein is provided by Sierra Analytics HDExaminer software. This is the most-widely used vendor-neutral solution available on the market and has a high degree of functionality and is easy-to-use.

 

Watch the video on Sierra Analytics HDExaminer software to find out how it can help you interpret the results of HDX-MS experiments: 

 

Visualization and Insights

The information and insights gained following data analysis can be used to probe protein structure conformational changes in the presence of a perturbation.  

 

The regions responsible for the changes can be readily visualized by linking to crystal structure visualization programs such as PyMol.

Crystal structure representations of the protein HtpG undergoing structural changes in the presence of ATP based on data from HDX-MS experiments.
Crystal structure representations of the protein HtpG undergoing structural changes in the presence of ATP based on data from HDX-MS experiments (click to enlarge)