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Rapid Determination of Antibody-Antigen Affinity by Mass Photometry
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Biochemistry
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JoVE Journal Biochemistry
Rapid Determination of Antibody-Antigen Affinity by Mass Photometry

Rapid Determination of Antibody-Antigen Affinity by Mass Photometry

DOI:
10.3791/61784-v

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10:17 min

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February 08, 2021

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,
1Biophysics Core Facility, National Heart, Lung, and Blood Institute,National Institutes of Health

Chapters

  • 00:05Introduction
  • 00:52Flow Chamber Preparation and Assembly
  • 02:02Antibody-Antigen Samples Preparation
  • 03:00Mass Photometry Measurement
  • 04:36Mass Photometry Data Analysis
  • 06:34Equilibrium Constant Value Calculation
  • 08:23Results: Representative Antibody-Antigen Affinity
  • 09:30Conclusion

Summary

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We describe a single-molecule approach to antigen-antibody affinity measurements using mass photometry (MP). The MP-based protocol is fast, accurate, uses a very small amount of material, and does not require protein modification.

Tags

Antibody-antigen AffinityMass PhotometryProtein Binding CharacterizationAntibody-antigen BindingProtein OligomerizationProtein-protein InteractionsAffinity MeasurementsRapid Binding AnalysisProtein Sample PreparationFlow Chamber SetupMicroscope Imaging

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