Peptide epitope-imprinted polymer microarrays for selective protein recognition. Application for SARS-CoV-2 RBD...

Area of Science:

• Biomaterials Science
• Analytical Chemistry
• Molecular Recognition

Background:

• Surface Plasmon Resonance Imaging (SPRi) enables label-free, multiplexed screening of binding kinetics.
• Molecularly Imprinted Polymers (MIPs) are synthetic receptors with tailored recognition capabilities.
• Developing robust platforms for rapid and specific viral protein detection is crucial.

Purpose of the Study:

• To establish a generic method for generating epitope-imprinted polymer microarrays on SPRi chips.
• To demonstrate the utility of these microarrays for synthesizing and screening high-affinity binders.
• To develop MIPs for recognizing the SARS-CoV-2 spike protein receptor binding domain (RBD).

Main Methods:

• Microspotting of template peptides (RBD nonapeptide GFNCYFPLQ) onto gold SPRi chips.
• Electrosynthesis of a polyscopoletin nanofilm to create one-step MIP arrays.
• Multiplexed, label-free screening of binding kinetics using SPRi.

Main Results:

• Achieved MIPs with dissociation constants (Kd) in the low nanomolar range for RBD, surpassing natural binding affinity.
• Demonstrated high affinity binding to SARS-CoV-2 virus-like particles with excellent discrimination against influenza A (H3N2) virus.
• Confirmed the critical role of specific amino acid sequences in template peptides for MIP binding specificity.

Conclusions:

• The developed MIP-based SPRi microarray platform is a versatile tool for both synthetic and screening applications.
• The MIPs exhibit superior affinity and specificity for SARS-CoV-2 RBD and viral particles.
• This approach offers a promising strategy for rapid diagnostics and molecular recognition applications.