Generation of dengue virus non-structural protein 1 antibodies using phage display and hybridoma technology
Tan, Li Xuan
Date of Issue2016-05-16
Duke NUS, Emerging Infectious Disease Programme
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.
Final Year Project (FYP)
Nanyang Technological University