The COVID-19 pandemic demonstrates that the emergence of novel human-pathogenic viruses can pose a challenge for global health. So far, over 765 million confirmed cases of COVID-19 have been reported to the World Health Organization, including more than 6,9 million deaths (May 2023). Consequently, it has been recognized that there is an urgent need to enhance pandemic preparedness, which includes research on novel technologies for the rapid generation of antiviral drugs and vaccines.
Monoclonal antibodies with therapeutic or prophylactic potential are potent tools against (emerging) viruses like SARS-CoV-2. Amongst these antibodies, broadly neutralizing antibodies (bnABs) are of special interest as they remain effective even if the virus mutates at a fast pace. Multiple methods for the generation of monoclonal antibodies are available to this date. However, in the light of pandemic preparedness, it is required to develop novel and optimize current methods for the accelerated production of antiviral monoclonal antibodies.
The aim of this project is to compare modern fluorescence-activated cell sorting & single cell sequencing-based approaches to the classical hybridoma technology for the generation of monoclonal antibodies. Optimized and streamlined protocols could decrease the development time by several weeks. Accordingly, the generated monoclonal antibodies would be available to the clinics earlier.
Here, we aim to establish the use of antibody humanized TRIANNI mice for the rapid generation of fully human monoclonal antibodies. For example, SARS-CoV-2 Spike receptor binding domain (RBD)-specific Memory B Cells (MBCs) will be isolated from splenocytes of immunized animals by fluorescence-activated cell sorting after prime and boost immunizations with SARS-CoV-2 Spike DNA. In single cell reactions, a cDNA library for the B cell receptor (BCR) genes of these antigen-specific MBCs will be constructed and sequenced. Based on bioinformatic analyses, selected BCR heavy and light chain genes will be synthesized and cloned into expression vectors in order to produce recombinant fully human monoclonal antibodies in cell culture. These antibodies will be characterized further and be tested e.g., for their neutralization capacity to identify potential bnABs.
Furthermore, it is of importance that optimized methods for the generation of monoclonal antibodies are applicable to a broad diversity of viruses. In Addition to SARS-CoV-2, other viruses like tick-borne encephalitis virus (Flaviviridae) and BK virus (Polyomaviridae) will be targets for the generation of monoclonal antibodies, since in both cases there are only insufficient options for prevention and treatment available.
In conclusion, this project will provide optimized processes for the rapid development of fully human monoclonal antibodies and therefore contribute to enhance pandemic preparedness, while simultaneously generating fully human monoclonal antibodies against multiple viruses.
