Generation, characterisation and application of SARS-CoV-2 protein antigens for COVID-19 rapid diagnostic purposes in the hospital and community

We aim to generate a range of stably transfected cell lines to help meet the demand for quality-verified recombinant SARS-CoV-2 structural proteins, which are desperately needed, and which we will use, to help in the development of diagnostics and vaccines. In so-doing our project will complement and expand the repertoire of proteins available on the COVID-19 Protein Portal. The platforms we establish will also be perfectly positioned for the rapid generation of other targets in the future. We will produce these in two different mammalian expression systems, CHO and HEK293 cells, characterise these by mass spectrometry and undertake glycosylation analysis of the Spike glycoprotein. In producing non-glycosylated forms of the protein for comparison, we will define the importance of this post-translational modification for folding, assembly of the trimer form and the ability of the protein to detect anti-spike antibodies from recovered COVID-19 patients. The recombinant authentic and characterised forms of the SARS-CoV-2 proteins will be assessed in diagnostic assays in development with our industrial partner Mologic, and in ELISA assays in the East Kent Hospital Trust at Ashford to profile antibody responses across recovered COVID-19 patients. The proposed research will address the WHO 'coordinated global research roadmap 2019 Novel Coronavirus Global Research and Innovation forum' that stipulates the need for reagents that support the development of 'reliable serological testing as well as assays that monitor response and midterm support development of diagnostic products to improve clinical processes to contribute to control/point of care' and for development of vaccines.