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生物化学
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Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization
JoVE 杂志
生物化学
This content is Free Access.
JoVE 杂志 生物化学
Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization

DOI:
10.3791/59408-v

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10:41 min

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April 05, 2019

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, , ,
1Wolfson Centre for Applied Structural Biology, The Alexander Silberman Institute of Life Science,The Hebrew University of Jerusalem, 2Wyatt Technology Corporation, 3Danyel Biotech Ltd

Chapters

  • 00:04Title
  • 00:50Preparation of the Buffer
  • 02:10IEX-MALS Experiment
  • 05:38Analysis of IEX-MALS Experimental Data
  • 08:27Results: Optimization of IEX-MALS Experiment for BSA
  • 10:02Conclusion

Summary

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This protocol describes the use of high-specificity ion-exchange chromatography with multi-angle light scattering for an accurate molar mass determination of proteins, protein complexes, and peptides in a heterogeneous sample. This method is valuable for quality assessment, as well as for the characterization of native oligomers, charge variants, and mixed-protein samples.

Tags

Ion Exchange ChromatographyIEXMulti-angle Light ScatteringMALSProtein SeparationProtein CharacterizationProtein Quality ControlOligomeric StateHomogeneityIdentityConformationStructurePost-translation ModificationsAnion ExchangeAIEXCation ExchangeCIEXBSABufferFPLCDLS

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