8.1 Biochemistry: A Polyvalent, Allosteric Whole-Virus Binding Platform for Norovirus Detection
Abstract
The objective of the research project is to synthesize and optimize a polyvalent DNA origami aptamer-displaying "claw" nanostructure that binds Norovirus with high selectivity and efficiency. The research effort will encompass three aims: I) Improving 1: 1 claw:virus binding selectivity and affinity in solution In prior work, claw nanostructures were assembled that exhibited selective binding to virus capsids but multimerized in solution, leading to several claw structures binding the same viral capsid molecule. This aim will screen various tethered/blocked claw designs to improve 1:1 capsid solution-phase binding efficiency and selectivity. Gel shift with FRET-labeled claws will be used to determine binding constants, allosteric response and binding specificities. 2) Claw surface isolation to obtain l: 1 binding/FRET analysis In parallel to Aim I , a surface-based assay system will be used to isolate claw nanostructures on a surface so they cannot diffuse, and thus cannot multimerize. Claw structures will be chemically anchored to glass surfaces displaying NHS-esters, streptavidin or functionalized alkenes. Binding of virus capsids will be analyzed using FRET. 3) Adapting Norovirus aptamers for polyvalent, serotype-selective binding This aim will focus on screening aptamers for binding to norovirus capsids using gel shift analysis. Selective aptamers will then be incorporated into the most successful claw nanostructure designs from Aims I and 2, and binding selectivity will be analyzed. Optimize claw aptamer recognition sites (before and after attachment to claw) to optimize binding strength and selectivity between different norovirus serotypes.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jan 12, 2017
- Source ID
- W911NF1610178
Entities
People
- Philip S. Lukeman
Organizations
- Army Contracting Command
- United States Army