A framework for automated multimodal HDX-MS analysis

We present pyHXExpress, a customizable codebase for automated high-throughput multimodal analysis of all spectra generated from HDX-MS experiments. The workflow was validated against a synthetic test dataset to test the fitting algorithms and to confirm the statistical outputs. We further establish a framework for the determination of multimodality throughout a protein system by rigorous evaluation of multimodal fits across all peptide spectra. We demonstrate this approach using entire protein datasets to detect multimodality, conformational heterogeneity, and characterize dynamics of small heat shock protein HSPB5 and two disease mutants.