Development of nanosensor photonic systems for rapid virus detection using directed evolution of protein platforms: the case of SARS-CoV-2 (BioPickMol)
Call: OPII-VA/DP/2020/9. -01
Type of action: Operational Program Integrated Infrastructure of the Slovak Republic – 311000
Proposal number: NFPP313010AUW6
Duration: 2021-2023
Partner: University of P. J. Safarik, Kosice, Slovakia
Abstract: The aim of the project was to create a unique technology for detecting the SARS-CoV-2 virus (with the prospect of using this technology to detect other types of viruses), which, compared to current methodologies, is characterized by a significantly higher level of sensitivity, selectivity, speed (several minutes) and also the possibility of performing in-situ analysis (in the field without the need to send samples to a specialized laboratory). The proposed technology is based on the selective binding of the virus to a nanostructured photonic chip through the functionalization of the surface of this chip with selected specific proteins or peptide linkers, which will be developed by the method of directed evolution of selected protein platforms or mathematical modeling. The subsequent detection of the presence of the virus is based on PickMol technology and thus the surface amplification of the Raman signal originating from the detected molecules through light-generated plasmons.
Result: Within the framework of the project, SAFTRA photonics developed a methodology for detecting selected SARS-Cov-2 variants differing in the structure of the binding domain of the Spike proteins (Spike RBD wild type, Spike RBD UK-variant (N501Y), Spike RBD South Africa-variant (K417N, E484K, N501Y), Spike RBD Brazil-variant (K417T, E484K, N501Y), Spike RBD India-variant (K452R, E484Q, P681R)). The specific and strong binding of these types of spike proteins to hACE2 cell receptors is ensured by modeling and subsequently synthesizing specific peptide linkers (consisting of 19 amino acids) imitating the binding domain of the hACE2 receptor of cells. The linkers were connected to the metallic structures of the PickMol nanostructure chip through a specific bond. Subsequently, SERS spectra of complexes of SARS-Cov-2 virus with peptide linkers formed on a nanostructured chip confirmed the unique capabilities of PickMol technology for sensitive, selective and rapid detection of various variants of SARS-Cov-2 viruses.
