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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
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A Competent Hepatocyte Model Examining Hepatitis B Virus Entry through Sodium Taurocholate Cotransporting Polypeptide as a Therapeutic Target

Article DOI: 10.3791/63761-v • 11:34 min • May 10th, 2022
May 10th, 2022 •
1, 2, 2, 3,4, 5
1Department of Biochemistry, Faculty of Pharmacy, Mahidol University, 2Program in Translational Medicine, Faculty of Medicine Ramathibodi Hospital, Mahidol University, 3Excellent Center for Drug Discovery, Faculty of Science, Mahidol University, 4Department of Biotechnology, Faculty of Science, Mahidol University, 5Department of Pharmacology, Faculty of Medicine Siriraj Hospital, Mahidol University

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Summary
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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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Hepatocyte Model Hepatitis B Virus Sodium Taurocholate Cotransporting Polypeptide Therapeutic Target Anti HBV Compound Viral Entry NTCP Binding Candidate Compound Measurement Of NTCP EDTA Ethylenediaminetetraacetic Acid Fix Hepatocytes Paraformaldehyde 1.5 Milliliter Microcentrifuge Tube Centrifugation Wash Cells Fetal Bovine Serum Permeabilize Cells Triton X 100 Primary Antibody Against NTCP Perm Wash Buffer
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