Master of Chemical Sciences Capstone Projects
Date of this Version
8-4-2020
Document Type
Capstone Report
Abstract
Hepatitis B virus (HBV) chronically infects more than 250 million people worldwide and causes 680, 000 deaths annually due to liver failure and hepatocellular carcinoma (HCC). HBV core protein allosteric modulators (CpAMs) disrupt HBV capsid assembly by interacting with the hydrophobic interface of core protein (Cp) dimers, the building block of HBV capsid. This project intended to establish an in vivo capsid assembly assay, to further understand the mechanism of HBV capsid assembly through interaction between CpAMs and Cp dimer. To this end, the capsid assembly in E. coli and mammalian cells were studied through native agarose gel electrophoresis-based particle gel assay. The results emphasize the necessity to dismiss confounding effects. C-terminally His-tagged wild-type HBV Cp were successfully overexpressed in E. coli, and purified through affinity column chromatography.
Images
Figure 6. Particle gel assay of capsids assembled in HepG2 cells treated with ENAN34017.
Figure 16. Validation of C-terminally His-tagged Cp1-149 and Cp1-183 protein capsid assembly by particle gel assay (PC: AML12 HBV polymerase Y63F cell lysate from Zhanying, NC: 1mg/mL lysozyme)
Keywords
HBV, capsid assembly, CpAM, protein expression
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 License.
Date Posted: 26 January 2021
Comments
At the time of publication, Shitian Wei was affiliated with Nanjing University. Currently, she is a Master student at the Department of Chemistry at the University of Pennsylvania.