Cross-Reactive Synbody Affinity Ligands for Capturing Diverse Noroviruses

Area of Science:

• Virology
• Immunology
• Biotechnology

Background:

• Noroviruses cause widespread acute gastroenteritis.
• Viral evolution necessitates novel detection methods and affinity ligands.
• Existing detection methods face challenges due to norovirus diversity.

Purpose of the Study:

• Develop novel affinity ligands for norovirus detection.
• Utilize synbody technology for broad norovirus strain recognition.
• Assess the potential of synbodies for diagnostic and therapeutic applications.

Main Methods:

• Generation of bivalent synbody affinity ligands using norovirus virus-like particles (VLPs).
• Characterization of synbody binding affinities (dissociation constants, K_D) against various norovirus genotypes.
• Evaluation of synbody performance in capturing norovirus from diluted clinical samples.

Main Results:

• Over 20 synbodies developed with low nanomolar dissociation constants (< 10 nM) for GII.4 VLP.
• Four synbodies demonstrated broad cross-reactivity with affinities ranging from 0.5 to 8 nM against multiple GI and GII genotypes.
• One synbody successfully captured GII.4 norovirus from a 200,000-fold dilution of a positive stool sample.
• Synbodies recognized and captured four different norovirus genotypes.

Conclusions:

• The synbody approach is effective in generating diverse norovirus affinity ligands.
• Developed synbodies show broad reactivity and high sensitivity for norovirus detection.
• Synbodies hold significant potential for future norovirus diagnostics and therapeutic development.