Dengue Virus prM-Specific Human Monoclonal Antibodies with Virus Replication-Enhancing Properties Recognize a Single Immunodominant Antigenic Site

Smith S, et al., 90(2):780-9, J Virol, 2015

Dengue is a mosquito-borne disease caused by four dengue virus (DENV) serotypes (DENV1 to DENV4). Currently there are no specific dengue therapeutics. It is crucial to have a better understanding of the pathogenesis of severe dengue in order to develop DENV vaccines. It is believed that cross-reactive antibodies produced during a primary infection play an important role in the pathogenesis of severe dengue. Described herein is the isolation of a large panel of dengue premembrane (prM) protein-specific human monoclonal antibodies (MAbs) from post-infected individuals in order to explore antigenic sites on the prM protein that is recognized by human antibodies. All competition (binding) assays were performed by Bio-Layer Interferometry(BLI) using a Pall ForteBio Octet RED instrument equipped with Streptavidin Biosensor probes. Biotinylated mouse anti-dengue virus prM MAb 2H2 was immobilized onto Biosensor tips. The sensor tips were then immersed in a solution of crude DENV2 S-16803 viron particles. After a washing step, the first human anti-DENV MAb was introduced. Then exposed to a second human MAb, in order to study binding interference. Results obtained from competition binding assays demonstrate that only a single antibody molecule can be bound to each prM protein molecule at any given time. Overall, the findings of this study indicate a distinct overlapping epitope that lies within a single major antigenic site. This antigenic domain was also suggested to represent an immunodominant site of the prM protein.

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