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A Competent Hepatocyte Model Examining Hepatitis B Virus Entry through Sodium Taurocholate Cotransporting Polypeptide as a Therapeutic Target
JoVE Journal
Biochemistry
Author Produced
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JoVE Journal Biochemistry
A Competent Hepatocyte Model Examining Hepatitis B Virus Entry through Sodium Taurocholate Cotransporting Polypeptide as a Therapeutic Target
DOI:

11:34 min

May 10, 2022

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Chapters

  • 00:05Introduction
  • 00:50Measuring the Sodium Taurocholate Co-Transporting Polypeptide (NTCP) Using Flow Cytometry
  • 04:30Anti-HBV (Hepatitis B Virus) Entry Assay
  • 06:13HBV (Hepatitis B Virus) -Cell Binding Assay
  • 06:50Taurocholic Acid Uptake Assay
  • 07:34Determination of Protein-Ligand Interactions Using Isothermal Titration Calorimetry
  • 09:14Results: Screening of Anti-Hepatitis B Virus Using Isothermal Titration Calorimetry
  • 10:36Conclusion

Summary

Automatic Translation

We present a protocol to screen anti-hepatitis B virus (HBV) compounds targeting pre- and post-viral entry lifecycle stages, using isothermal titration calorimetry to measure binding affinity (KD) with host sodium taurocholate cotransporting polypeptide. Antiviral efficacy was determined through the suppression of viral lifecycle markers (cccDNA formation, transcription, and viral assembly).

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