A pair of non-competing neutralizing human monoclonal antibodies protecting from disease in a SARS-CoV-2 infection model

Peter, Antonia Sophia; Roth, Edith; Schulz, Sebastian R.; Fraedrich, Kirsten; Steinmetz, Tobit; Damm, Dominik; Hauke, Manuela; Richel, Elie; Mueller-Schmucker, Sandra; Habenicht, Katharina; Eberlein, Valentina; Issmail, Leila; Uhlig, Nadja; Dolles, Simon; Grüner, Eva; Peterhoff, David; Ciesek, Sandra; Hoffmann, Markus; Pöhlmann, Stefan; McKay, Paul F.; Shattock, Robin J.; Wölfel, Roman; Wagner, Ralf; Eichler, Jutta; Schuh, Wolfgang; Neipel, Frank; Ensser, Armin; Mielenz, Dirk; Tenbusch, Matthias; Winkler, Thomas H.; Grunwald, Thomas; Überla, Klaus; Jäck, Hans-Martin

TRIANNI mice carry an entire set of human immunoglobulin V region gene segments and are a powerful tool to rapidly generate human monoclonal antibodies. After immunizing these mice against the spike protein of SARS-CoV-2, we identified 29 hybridoma antibodies that reacted with the SARS-CoV-2 spike protein. Nine antibodies neutralized SARS-CoV-2 infection at IC50 values in the subnanomolar range. ELISA-binding studies and DNA sequence analyses revealed one cluster of clonally related neutralizing antibodies that target the receptor-binding domain and compete with the cellular receptor hACE2. A second cluster of neutralizing antibodies binds to the N-terminal domain of the spike protein without competing with the binding of hACE2 or cluster 1 antibodies. SARS-CoV-2 mutants selected for resistance to an antibody from one cluster are still neutralized by an antibody from the other cluster. Antibodies from both clusters markedly reduced viral spread in mice transgenic for human ACE2 and protected the animals from SARS-CoV-2 induced weight loss. Thus, we report two clusters of potent non-competing SARS-CoV-2 neutralizing antibodies providing potential candidates for therapy and prophylaxis of COVID-19. The study further supports the use of transgenic animals with human immunoglobulin gene repertoires in pandemic preparedness initiatives.