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Collection of Monoclonal Antibodies Targeting SARS-CoV-2 Proteins
Marina Pribanić Matešić1, Paola Kučan Brlić1, Tihana Lenac Roviš1
1Center for Proteomics, Faculty of Medicine, University of Rijeka, Braće Branchetta 20, 51000 Rijeka, Croatia.
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.
Insights
Researchers developed over 40 new monoclonal antibodies (mAbs) targeting various SARS-CoV-2 proteins. These specific molecular tools aid in understanding viral pathogenesis and designing new COVID-19 therapies.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- The COVID-19 pandemic necessitates advanced research tools for understanding SARS-CoV-2.
- High-quality molecular tools, such as monoclonal antibodies, are crucial for scientific advancement.
Purpose of the Study:
- To develop and characterize a novel collection of monoclonal antibodies against SARS-CoV-2 proteins.
- To provide specific reagents for comprehensive analysis of the SARS-CoV-2 proteome.
Main Methods:
- Expression and purification of recombinant SARS-CoV-2 proteins for use as immunogens.
- Development of hybridomas and screening of monoclonal antibody (mAb) clones.
- Testing mAb binding to recombinant proteins and infected cells using flow cytometry, immunofluorescence, and Western blot.
Main Results:
- Generation of over 40 new monoclonal antibodies against SARS-CoV-2.
- mAbs were produced against structural proteins (Spike, Nucleocapsid) and non-structural proteins (nsp1, nsp7-10, nsp16) and accessory factors (ORF3a, ORF9b).
- The developed mAbs demonstrated high specificity and applicability in various detection methods.
Conclusions:
- The new collection of monoclonal antibodies offers valuable tools for SARS-CoV-2 research.
- These antibodies will facilitate a deeper understanding of viral pathogenesis.
- The mAbs can aid in the design and development of therapeutic strategies against COVID-19.
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