Semi-automated data processing of hydrogen exchange mass spectra using HX-Express

Abstract

A Microsoft Excel utility, HX-Express, that significantly accelerates the analysis of hydrogen exchange mass spectrometry data is described. HX-Express generates deuterium uptake and peak width plots from peaks in mass spectral data. Data analysis is intentionally semi-automated, requiring that the user find the peaks to be analyzed. The peaks are entered in the form of x, y lists of m/z versus intensity or can be directly imported from Waters MassLynx software. Analysis of data with HX-Express provides the same results as manual data processing but is substantially faster. In addition to speed, HX-Express provides and preserves visual and numeric displays of the analysis process for quality control.

Binding of Cyclic Diguanylate in the Non-catalytic EAL Domain of FimX Induces a Long-range Conformational Change

“…The time from initiation of digestion to elution of the last peptide was ϳ25 min. Xcalibur software was used for spectrum analysis and data extraction, and the H/D exchange data were processed using the program HX-Express (27). Measured peptide masses were corrected for artifactual in-exchange and back-exchange following the methods described by Hoofnagle et al (23).…”

Recognition of N-Glycoforms in Human Chorionic Gonadotropin by Monoclonal Antibodies and Their Interaction Motifs

“…The experimental monoisotopic m/z values corresponding to all peptides in unexchanged form were input into the database to look for matches. The deuterium levels of each assigned peptides were then determined using the HX-Express software (27).…”

HDX Match Software for the Data Analysis of Top-Down ECD-FTMS Hydrogen/Deuterium Exchange Experiments

“…Numerous approaches have been developed mainly for the processing of bottom-up HDX data, ranging from simple peak processing utilities [4] to fully integrated instrument-software platform solutions [3,[5][6][7]. Arguably, the most advanced approaches for determining deuteration levels combine the identification of the deuterated peptide (thereby determining its elemental composition), calculation of its deuterated isotopic distribution, and fitting it to the experimental data.…”