A Decoupled Automation Platform for Hydrogen/Deuterium Exchange Mass Spectrometry Experiments
- 13 November 2019
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Society for Mass Spectrometry
- Vol. 30 (12), 2580-2583
- https://doi.org/10.1007/s13361-019-02331-2
Abstract
Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a biophysical technique well suited to the characterization of protein dynamics and protein–ligand interactions. In order to accurately define the rate of exchange, HDX experiments require the repeated measure of deuterium incorporation into the target protein across a range of time points. Accordingly, the HDX-MS experiment is well suited to automation, and a number of automated systems for HDX-MS have been developed. The most widely utilized platforms all operate an integrated design, where robotic liquid handling is interfaced directly with a mass spectrometer. With integrated designs, the exchange samples are prepared and injected into the LC-MS following a “real-time” serial workflow. Here we describe a new HDX-MS platform that is comprised of two complementary pieces of automation that disconnect the sample preparation from the LC-MS analysis. For preparation, a plate-based automation system is used to prepare samples in parallel, followed by immediate freezing and storage. A second piece of automation has been constructed to perform the thawing and LC-MS analysis of frozen samples in a serial mode and has been optimized to maximize the duty cycle of the mass spectrometer. The decoupled configuration described here reduces experiment time, significantly improves capacity, and improves the flexibility of the platform when compared with a fully integrated system.Keywords
This publication has 19 references indexed in Scilit:
- Interlaboratory Comparison of Hydrogen–Deuterium Exchange Mass Spectrometry Measurements of the Fab Fragment of NISTmAbAnalytical Chemistry, 2019
- Integrating biophysics with HTS-driven drug discovery projectsDrug Discovery Today, 2016
- A Binding Site on IL-17A for Inhibitory Macrocycles Revealed by Hydrogen/Deuterium Exchange Mass SpectrometryJournal of Medicinal Chemistry, 2016
- HDX reveals unique fragment ligands for the vitamin D receptorBioorganic & Medicinal Chemistry Letters, 2014
- HDX Workbench: Software for the Analysis of H/D Exchange MS DataJournal of the American Society for Mass Spectrometry, 2012
- Probing protein interactions with hydrogen/deuterium exchange and mass spectrometry—A reviewAnalytica Chimica Acta, 2012
- Hydrophobic Interactions Improve Selectivity to ERα for Benzothiophene SERMsACS Medicinal Chemistry Letters, 2012
- DNA binding alters coactivator interaction surfaces of the intact VDR–RXR complexNature Structural & Molecular Biology, 2011
- Unique Ligand Binding Patterns between Estrogen Receptor α and β Revealed by Hydrogen−Deuterium ExchangeBiochemistry, 2009
- The Deuterator: software for the determination of backbone amide deuterium levels from H/D exchange MS dataBMC Bioinformatics, 2007