Optimising expression and purification of Dengue soluble E protein

• Main Author: Poh, Nina Yong Xin
• Other Authors: Xiao Tianshu
• Format: Final Year Project
• Language: English
• Published: Nanyang Technological University 2023
• Subjects: Science::Biological sciences
• Online Access: https://hdl.handle.net/10356/169794
• Institution: Nanyang Technological University

This study explores the significance of expression and purification of soluble dengue Envelope (E) protein which has strong applications in viral biology, vaccine development, and therapeutic strategies. Dengue, a type of flavivirus, is a widespread mosquito-borne infection lacking effective antiviral treatments. The E protein is crucial for viral fusion and host cell entry. The soluble E (sE) protein, obtained by truncation of Transmembrane Domain (TMD), serves as an antigen for serological assays, aiding diagnosis and seroprevalence assessment. It offers insights into viral-host immune interactions. The full-length E protein is essential for vaccine design. The study aims to express and purify the soluble E protein, modified with Tobacco Etch Virus (TEV) and Twin-Strep-tag®, for applications such as structural study and high throughput screening. Successful modification and vector expression in pcDNA3.1 enhance solubility, enabling drug development and testing. This project sets a baseline for future functional and mechanistic understanding. Overall, it advances understanding of Dengue virus biology and potential therapeutic advances.