3 years ago

Cross-Reactive Synbody Affinity Ligands for Capturing Diverse Noroviruses

Cross-Reactive Synbody Affinity Ligands for Capturing Diverse Noroviruses
Stephen Albert Johnston, Hugh S. Mason, Luhui Shen, John C. Lainson, Chris W. Diehnelt, Paul E. Belcher, Nidhi Gupta
Noroviruses are the most common cause of acute gastroenteritis in the developed world. Noroviruses are a diverse group of nonenveloped RNA viruses that are continuously evolving. This leads to the rise of immunologically distinct strains of the same genotype on a frequent basis. This diversity presents a unique challenge for detection and tracking of new strains, with the continuous need for new norovirus affinity ligands. Our group developed a family of bivalent synbody affinity ligands using a virus-like particle (VLP) from the 2006 GII.4 Minerva strain of norovirus. We produced more than 20 synbodies with low nanomolar dissociation constants (KD < 10 nM) for GII.4 VLP. We measured binding affinity for four synbodies against VLPs from multiple GI and GII genotypes and found that the synbodies were broadly cross-reactive with affinities that ranged from 0.5 to 8 nM. We tested the ability of these synbodies to capture norovirus from dilute solutions and found that one synbody could capture GII.4 from a 200 000-fold dilution from a norovirus positive stool sample. When these synbodies were tested for the ability to capture of multiple genotypes, we found that four different genotypes were recognized. These data demonstrate that the synbody approach can generate multiple affinity ligands for future use in norovirus detection and possible therapeutic development.

Publisher URL: http://dx.doi.org/10.1021/acs.analchem.7b01337

DOI: 10.1021/acs.analchem.7b01337

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