Biochemical Adjustments of native EBOV Glycoprotein in Patient Sample to Unmask target-Epitopes for Rapid Diagnostic Testing
Early detection is important for mobilizing swift response and control. Existing technologies for filovirus detection are not suited for point of care (POC) use, being expensive, not fast-enough and requiring laboratory facilities absent in many villages where index cases occur. While 2 rapid diagnostic tests (RDTs) have recently emerged detecting EBOV at the point of care (POC), there are no pan-filovirus targeted RDTs. Pan-filovirus RDTs are required to ensure early detection, response and control of the on-going and future outbreaks. Moreover, the mAbs presented are candidate therapeutics.
This project could yield the 1st ever prototypes of RDTs for the duo-detection of EBOV and MARV.
Post-translational modifications, including secondary structure folding, glycosylation (with mannose glycan residues) and or disulfide bonding of the GP1 and GP2 sub-units, might be obstructing mAb detection of native EBOV Gp by the sandwich EIA. The aim is to exhaustively study the 3-D crystal structure of EBOV GP (PDB entry #3CSY) for these changes and perform computational remodeling to predict impact of treatment with biochemical reagents that remove the post-translational modifications unveiled.
To apply either the glycosidases, endopeptidases, and reducing agents in sample buffer, we must optimize their concentrations. Optimizing glycosidase, endopeptidase or reducing agent concentrations for use as sample buffer for duo-filovirus antigen detection. Analyses of the results to determine optimal concentrations of glycosidase, endopeptidase, or reducing agent for use as sample buffer for duo-filovirus antigen detection
This project could yield the 1st ever prototypes of RDTs for the duo-detection of EBOV, other ebolavirus spp and MARV. Pan-filovirus RDTs are required to ensure early detection, response and control of the on-going and future outbreaks. Specifically, we are looking at hastening the validation of an existing technology to enable its deployment and use in the response geared at controlling and containing EVD outbreaks. Our antigen and host-specific IgM detecting RDTs are relevant towards diagnosing acute EBOV infection, while IgG RDTs and or assays might be relevant towards epidemiological evaluation for prior exposure to EBOV, or assessing the extent of vaccine immunological responses.
The AdjustEBOVGP-Dx team consist of academics with acknowledged expertise in drug design and antiviral research.
SLU, Uppsala UniversitySweeden
Agricultural University of Athens,Greece
Warsaw University of Technology,Polland
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