Generation of antibodies to assess specific dengue immune memory
Lee, Bernie Yi Jie
Date of Issue2017-04-25
School of Biological Sciences
Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR)
Almost half of the global population is at risk of dengue virus (DENV) infection. Due to the presence of four different DENV serotypes, elucidating the complex interaction of our immune system against DENV has been challenging. To understand the mechanisms of protective immunity against DENV, B-cell immune responses are commonly examined via ELISpot assay or antibodies characterization. However, ELISpot assay is restricted to the analysis of one dengue serotype per well. Moreover, the low efficiency of B-cells immortalization by Epstein-Barr virus during antibodies production may impede elucidating the precise repertoire of B-cell immune response against dengue infections. The development of a Quad-Color FluoroSpot (QCF) assay can address limitations faced in current assays and assess for serotype-specific dengue immune memory. Thus, serotype-specific antibodies are required as detection tools. This study explored utilizing fluorescently labelled DENV as antigen-baits in flow cytometry to isolate DENV-specific memory B-cells that may potentially be serotype-specific. A total of 28 antibodies were generated and their binding specificities towards DENV were characterized. Future optimization of such strategy in flow cytometry could possibly allow for the isolation and expression of rare DENV serotype-specific mAbs that can be used as detection tools to facilitate the development of the QCF assay.
Final Year Project (FYP)
Nanyang Technological University