Abstract
In early 2020, the COVID-19 pandemic sparked a global crisis that continues to pose a serious threat to human health and the economy. Further advancement in research is necessary and requires the availability of quality molecular tools, including monoclonal antibodies. Here, we present the development and characterization of a collection of over 40 new monoclonal antibodies directed against different SARS-CoV-2 proteins. Recombinant SARS-CoV-2 proteins were expressed, purified, and used as immunogens. Upon development of specific hybridomas, the obtained monoclonal antibody (mAb) clones were tested for binding to recombinant proteins and infected cells. We generated mAbs against structural proteins, the Spike and Nucleocapsid protein, several non-structural proteins (nsp1, nsp7, nsp8, nsp9, nsp10, nsp16) and accessory factors (ORF3a, ORF9b) applicable in flow cytometry, immunofluorescence, or Western blot. Our collection of mAbs provides a set of novel, highly specific tools that will allow a comprehensive analysis of the viral proteome, which will allow further understanding of SARS-CoV-2 pathogenesis and the design of therapeutic strategies.
Highlights
In December of 2019, an outbreak of a previously unknown coronavirus began inWuhan, China
Our work provides a collection of monoclonal antibodies against different SARS-CoV-2 proteins that will allow a comprehensive analysis of viral proteins and provide tools to systematically investigate viral pathogenesis in the future studies
Analysis of antibodies raised against ORF3a showed binding to the ORF3a peptide conjugated to the bovine serum albumin (BSA) carrier, and protein band corresponding to the molecular weight of ORF3a could not be detected
Summary
In December of 2019, an outbreak of a previously unknown coronavirus began inWuhan, China. COVID-19, has spread rapidly throughout the world, and the World Health Organization declared a pandemic of this novel virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), on 11 March 2020. Its genome is a positive-sense, single-stranded RNA of ~30 kb in length which encodes 16 non-structural proteins, four structural proteins (Nucleocapsid, Spike, Envelope, and Membrane) and several accessory factors [4,5]. We report the development and characterization of over 40 SARS-CoV-2 monoclonal antibodies directed against structural proteins (Nucleocapsid and Spike), several non-structural proteins (nsp, nsp, nsp, nsp, nsp, nsp16) and accessory factors (ORF3a, ORF9b). Our work provides a collection of monoclonal antibodies against different SARS-CoV-2 proteins that will allow a comprehensive analysis of viral proteins and provide tools to systematically investigate viral pathogenesis in the future studies
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