HDX Solution

The HDX Workflow

HDX-MS experiments are often a series of time-course experiments performed under at least 2 different conditions e.g. at 2 different pH levels, or at 2 different temperatures, or with and without an additive. 

 

Once the conditions for the series of experiments have been ascertained, the information is transferred to the Chronos scheduling software for automatic calculation of the pre-incubation, labelling, quenching, digestion and separation times. 

Flowchart representing the tasks performed automatically by the HDX Solution for a complete HDX-MS experiment of HtpG/ HtpG+ATP
Flowchart representing the tasks performed automatically by the HDX Solution for a complete HDX-MS experiment of HtpG/ HtpG+ATP (click to enlarge)

Once everything is set-up and the acquisition started the system runs automatically preparing, injecting and analysing all the samples in the series.  After the mass spectrometric data has been collected this is uploaded to the HDExaminer software for calculation of the H/D ratios, generation of the deuteration uptake plots and production of heat maps. These data can then be exported to a 3-D crystal structure program such as PyMol for a 3-D visualization of the regions affected by the change being studied.
 

HDX Solution Workflow (click to enlarge):

HDX Solution Workflow
HDX Solution Workflow