Peptide-Integrated Superparamagnetic Nanoparticles for the Identification of Epitopes from SARS-CoV‑2 Spike and Nucleocapsid Proteins

The COVID-19 pandemic, caused by the fast transmission and spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is currently considered a serious health problem, requiring an effective strategy to contain SARS-CoV-2 dissemination. For this purpose, epitopes of the SARS-CoV-2 spike (S) and sucleocapsid (N) proteins were identified by bioinformatics tools, and peptides that mimic these epitopes were chemically synthesized and then conjugated to superparamagnetic nanoparticles (SPMNPs). Three peptides from S protein and three from N protein were used as antigens in a conventional enzyme-linked immunosorbent assay (ELISA) against serum samples from COVID-19-positive patients, or from healthy donors, collected before the pandemic. Three peptides were effective as antigens in conventional peptide-based ELISA, achieving 100% sensitivity and specificity, with high accuracy. The best-performing peptides, p2pS, p1pN, and p3pN, were associated with superparamagnetic nanoparticles (SPMNPs) and were used to perform nanomagnetic peptide-based ELISA. The p2pS–SPMNP conjugate presented 100% sensitivity and specificity and excellent accuracy (area under the curve (AUC) = 1.0). However, p1pN and p3pN peptides, when conjugated to SPMNPs, did not preserve the capacity to differentiate positive sera from negative sera in all tested samples, yet both presented sensitivity and specificity above 80% and high accuracy, AUC > 0.9. We obtained three peptides as advantageous antigens for serodiagnosis. These peptides, especially p2pS, showed promising results in a nanomagnetic peptide-based ELISA and may be suitable as a precoated antigen for commercial purposes, which would accelerate the diagnosis process.