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Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry
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Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry

Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry

DOI:
10.3791/62672-v

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05:45 min

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June 08, 2021

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, , , , ,
1School of Chemistry and Materials Science,Nanjing Normal University, 2Institute for Cell Analysis,Shenzhen Bay Laboratory, 3Department of Chemistry,University of British Columbia

Capitoli

  • 00:04Introduction
  • 00:38Operation of CE-Based HDX MS Analysis
  • 04:01Results: Conformational Characterization of Proteins with Top-Down Mass Spectrometry
  • 05:18Conclusion

Summary

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Presented here is a protocol for a capillary electrophoresis-based hydrogen/deuterium exchange (HDX) approach coupled with top-down mass spectrometry. This approach characterizes the difference in higher-order structures between different protein species, including proteins in different states and different proteoforms, by conducting concurrent differential HDX and electrophoretic separation.

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Keywords: Capillary ElectrophoresisHydrogen/deuterium ExchangeConformational CharacterizationTop-down Mass SpectrometryProtein Species SeparationHigher-order StructureHDX-CE-MSElectrophoretic EffectPreconditioningUltrasonicationBackground ElectrolyteInfusion TubingNano-electrospray IonizationInjection Volume EstimationElectrophoretic Voltage

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