The COVID-19 pandemic has exposed the extent of global connectivity and collective vulnerability to emerging diseases. From its suspected origins in Wuhan, China, it spread to all corners of the world in a matter of months. The absence of high-performance, rapid diagnostic methods that could identify asymptomatic carriers contributed to its worldwide transmission. Serological tests offer numerous benefits compared to other assay platforms to screen large populations. First-generation assays contain targets that represent proteins from SARS-CoV-2. While they could be quickly produced, each actually has a mixture of specific and non-specific epitopes that vary in their reactivity for antibodies. To generate the next generation of the assay, epitopes were identified in three SARS-Cov-2 proteins (S, N, and Orf3a) by SPOT synthesis analysis. After their similarity to other pathogen sequences was analyzed, 11 epitopes outside of the receptor-binding domain (RBD) of the spike protein that showed high reactivity and uniqueness to the virus. These were incorporated into a ß-barrel protein core to create a highly chimeric protein. Another de novo protein was designed that contained only epitopes in the RBD. In-house ELISAs suggest that both multiepitope proteins can serve as targets for high-performance diagnostic tests. Our approach to bioengineer chimeric proteins is highly amenable to other pathogens and immunological uses.
【저자키워드】 COVID-19, SARS-CoV-2, IgG, coronavirus, De novo design, linear B cell epitopes, serodiagnostic, 【초록키워드】 serological test, antibodies, Diseases, COVID-19 pandemic, Transmission, Asymptomatic carrier, virus, ELISA, Protein, pathogen, RBD, diagnostic tests, target, epitope, platform, non-specific, Diagnostic method, Analysis, similarity, chimeric, sequence, de novo, while, offer, approach, populations, immunological, benefit, Wuhan, China, produced, analyzed, identify, spread to, generate, absence, the spike protein, the RBD, the receptor-binding domain, contributed, reactivity, 【제목키워드】 detection, Region, RBD, the Spike,