Generation of dengue virus non-structural protein 1 antibodies using phage display and hybridoma technology

• Main Author: Tan, Li Xuan
• Other Authors: Subhash Vasudevan
• Format: Final Year Project
• Language: English
• Published: 2016
• Subjects: DRNTU::Science::Biological sciences
• Online Access: http://hdl.handle.net/10356/67372
• Institution: Nanyang Technological University

Dengue Virus (DENV) is an emerging flavivirus that is increasingly endemic worldwide with no efficacious antiviral treatments. Non-Structural Protein 1 (NS1) is an intriguing one that assumes diverse roles during infection ranging from a cofactor in viral replication complex to a disease aggravator. However, the functions of NS1 are not fully elucidated yet and antibodies would make a useful immunochemical tool to understand it better. Thus, NS1 antibodies are generated using phage display and hybridoma technology. In this project, a chimeric Fab-phage immunized library with a diversity of 4.828 x 10^6 cfu was constructed and screened for strong NS1 Fab-phage binders. Hybridomas produced were also screened for positive NS1 antibody hybridoma producers. As a result, 11 unique clones from phage display and a polyclonal hybridoma culture (2G12) were found. 2G12 is able to bind to NS1 in IFA and seems to bind to NS1 monomer in WB.