A Decoupled Automation Platform for Hydrogen/Deuterium Exchange Mass Spectrometry Experiments

Espada, Alfonso; Haro, Ruben; Castañon, Jesus; Sayago, Cristina Megías; Perez-Cozar, Francisco; Cano, Leticia; Redero, Pablo; Molina-Martin, Manuel; Broughton, Howard B.; Stites, Ryan E.; Pascal, Bruce D.; Griffin, Patrick R.; Dodge, Jeffrey A.; Chalmers, Michael J.

doi:10.1007/s13361-019-02331-2

Cited by 17 publications

(18 citation statements)

References 19 publications

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“…HDXMS experiments were performed using a decoupled system as previously described ( 47 ) with some modifications. Experiments were conducted at pH 7.4, in 10 mM sodium phosphate buffer, containing 150 mM NaCl (1x PBS buffer) in triplicate.…”

Section: Methodsmentioning

confidence: 99%

An anti-ANGPTL3/8 antibody decreases circulating triglycerides by binding to a LPL-inhibitory leucine zipper-like motif

Balasubramaniam¹,

Schroeder²,

Russell

et al. 2022

Journal of Lipid Research

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Section: Methodsmentioning

confidence: 99%

An anti-ANGPTL3/8 antibody decreases circulating triglycerides by binding to a LPL-inhibitory leucine zipper-like motif

Balasubramaniam¹,

Schroeder²,

Russell

et al. 2022

Journal of Lipid Research

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“…Lastly, we want to briefly bring attention to some articles that are important to HDX MS in their own right, which may guide and drive some of the future development of the field outside of the broad areas of LC, MS, or data processing. These articles include use of HDX MS in biopharma to monitor changes due to chemical modification (e.g., methionine oxidation), decoupled automation wherein sample preparation is a separate automation process from automated LC–MS, deamidation and protein stability, optimizing various points along the experimental workflow, including and exploiting PTMs during analysis, internal standards that report on the exchange conditions and back-exchange along the analysis pathway in order to correct for experimental condition fluctuations, capillary electrophoresis for separating coexisting protein states that are all deuterated together, and corrections to account for buffer additives that can alter the intrinsic rate of exchange …”

Section: Methods Development: Othermentioning

confidence: 99%

Developments in Hydrogen/Deuterium Exchange Mass Spectrometry

et al. 2020

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“…Soon, the very first HDX-UPLC system was commercialized by Waters (originally published by Engen [67]) and research groups also started repurposing pipettor robots for HDX sample preparation [31]. Recent research sponsored by Eli Lilly showed that complete automation is also possible via building a decoupled automation platform [13].…”

Section: Workflow and Equipmentmentioning

confidence: 99%

Hydrogen-Deuterium Exchange Mass Spectrometry: A Novel Structural Biology Approach to Structure, Dynamics and Interactions of Proteins and Their Complexes

Ozohanics

Ambrus

2020

Life

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Hydrogen/Deuterium eXchange Mass Spectrometry (HDX-MS) is a rapidly evolving technique for analyzing structural features and dynamic properties of proteins. It may stand alone or serve as a complementary method to cryo-electron-microscopy (EM) or other structural biology approaches. HDX-MS is capable of providing information on individual proteins as well as large protein complexes. Owing to recent methodological advancements and improving availability of instrumentation, HDX-MS is becoming a routine technique for some applications. When dealing with samples of low to medium complexity and sizes of less than 150 kDa, conformation and ligand interaction analyses by HDX-MS are already almost routine applications. This is also well supported by the rapid evolution of the computational (software) background that facilitates the analysis of the obtained experimental data. HDX-MS can cope at times with analytes that are difficult to tackle by any other approach. Large complexes like viral capsids as well as disordered proteins can also be analyzed by this method. HDX-MS has recently become an established tool in the drug discovery process and biopharmaceutical development, as it is now also capable of dissecting post-translational modifications and membrane proteins. This mini review provides the reader with an introduction to the technique and a brief overview of the most common applications. Furthermore, the most challenging likely applications, the analyses of glycosylated and membrane proteins, are also highlighted.

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A Decoupled Automation Platform for Hydrogen/Deuterium Exchange Mass Spectrometry Experiments

Cited by 17 publications

References 19 publications

An anti-ANGPTL3/8 antibody decreases circulating triglycerides by binding to a LPL-inhibitory leucine zipper-like motif

An anti-ANGPTL3/8 antibody decreases circulating triglycerides by binding to a LPL-inhibitory leucine zipper-like motif

Developments in Hydrogen/Deuterium Exchange Mass Spectrometry

Hydrogen-Deuterium Exchange Mass Spectrometry: A Novel Structural Biology Approach to Structure, Dynamics and Interactions of Proteins and Their Complexes

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