In Vitro Assembly and Stabilization of Dengue and Zika Virus Envelope Protein Homo-Dimers

In Vitro Assembly and Stabilization of Dengue and Zika Virus Envelope Protein Homo-Dimers

Blog Article

Abstract Zika virus (ZIKV) and the 4 dengue virus (DENV) serotypes are mosquito-borne Flaviviruses that are associated with severe neuronal and hemorrhagic syndromes.The mature flavivirus infectious virion has 90 envelope (E) protein homo-dimers that pack tightly to form a smooth protein coat with icosahedral symmetry.Human antibodies that strongly neutralize ZIKV and DENVs recognize complex quaternary structure epitopes displayed on E-homo-dimers and higher order structures.The ZIKV and DENV E car mackwell protein expressed as a soluble protein is mainly a monomer that does not display quaternary epitopes, which may explain the modest success with soluble recombinant E (sRecE) as a vaccine and diagnostic antigen.New strategies are needed to design recombinant immunogens that display these critical immune targets.

Here we present two novel methods for building or stabilizing in vitro E-protein homo-dimers that display quaternary epitopes.In the first approach we immobilize sRecE to enable subsequent dimer generation.As an alternate method, we describe the use of human mAbs to stabilize homo-dimers in solution.The ability to produce recombinant E protein dimers displaying quaternary structure epitopes is firestorm darts an important advance with applications in flavivirus diagnostics and vaccine development.