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Antiviral Res
. 2023 Dec 6:105778.
doi: 10.1016/j.antiviral.2023.105778. Online ahead of print. Nanobodies to multiple spike variants and inhalation of nanobody-containing aerosols neutralize SARS-CoV-2 in cell culture and hamsters
Metin Aksu 1 , Priya Kumar 2 , Thomas Güttler 3 , Waltraud Taxer 1 , Kathrin Gregor 1 , Bianka Mußil 1 , Oleh Rymarenko 1 , Kim M Stegmann 2 , Antje Dickmanns 2 , Sabrina Gerber 2 , Wencke Reineking 4 , Claudia Schulz 5 , Timo Henneck 6 , Ahmed Mohamed 6 , Gerhard Pohlmann 7 , Mehmet Ramazanoglu 7 , Kemal Mese 8 , Uwe Groß 8 , Tamar Ben-Yedidia 9 , Oded Ovadia 9 , Dalit Weinstein Fischer 9 , Merav Kamensky 9 , Amir Reichman 9 , Wolfgang Baumgärtner 4 , Maren von Köckritz-Blickwede 6 , Matthias Dobbelstein 10 , Dirk Görlich 11
Affiliations
The ongoing threat of COVID-19 has highlighted the need for effective prophylaxis and convenient therapies, especially for outpatient settings. We have previously developed highly potent single-domain (VHH) antibodies, also known as nanobodies, that target the Receptor Binding Domain (RBD) of the SARS-CoV-2 Spike protein and neutralize the Wuhan strain of the virus. In this study, we present a new generation of anti-RBD nanobodies with superior properties. The primary representative of this group, Re32D03, neutralizes Alpha to Delta as well as Omicron BA.2.75; other members neutralize, in addition, Omicron BA.1, BA.2, BA.4/5, and XBB.1. Crystal structures of RBD-nanobody complexes reveal how ACE2-binding is blocked and also explain the nanobodies' tolerance to immune escape mutations. Through the cryo-EM structure of the Ma16B06-BA.1 Spike complex, we demonstrated how a single nanobody molecule can neutralize a trimeric spike. We also describe a method for large-scale production of these nanobodies in Pichia pastoris, and for formulating them into aerosols. Exposing hamsters to these aerosols, before or even 24 h after infection with SARS-CoV-2, significantly reduced virus load, weight loss and pathogenicity. These results show the potential of aerosolized nanobodies for prophylaxis and therapy of coronavirus infections.
Keywords: Aerosol; Animal model; COVID-19; Hamster; Inhalation; Nanobodies; Protein structure; Receptor binding domain; SARS-CoV-2; Spike; VHH antibodies; Variants of concern.
. 2023 Dec 6:105778.
doi: 10.1016/j.antiviral.2023.105778. Online ahead of print. Nanobodies to multiple spike variants and inhalation of nanobody-containing aerosols neutralize SARS-CoV-2 in cell culture and hamsters
Metin Aksu 1 , Priya Kumar 2 , Thomas Güttler 3 , Waltraud Taxer 1 , Kathrin Gregor 1 , Bianka Mußil 1 , Oleh Rymarenko 1 , Kim M Stegmann 2 , Antje Dickmanns 2 , Sabrina Gerber 2 , Wencke Reineking 4 , Claudia Schulz 5 , Timo Henneck 6 , Ahmed Mohamed 6 , Gerhard Pohlmann 7 , Mehmet Ramazanoglu 7 , Kemal Mese 8 , Uwe Groß 8 , Tamar Ben-Yedidia 9 , Oded Ovadia 9 , Dalit Weinstein Fischer 9 , Merav Kamensky 9 , Amir Reichman 9 , Wolfgang Baumgärtner 4 , Maren von Köckritz-Blickwede 6 , Matthias Dobbelstein 10 , Dirk Görlich 11
Affiliations
- PMID: 38065245
- DOI: 10.1016/j.antiviral.2023.105778
The ongoing threat of COVID-19 has highlighted the need for effective prophylaxis and convenient therapies, especially for outpatient settings. We have previously developed highly potent single-domain (VHH) antibodies, also known as nanobodies, that target the Receptor Binding Domain (RBD) of the SARS-CoV-2 Spike protein and neutralize the Wuhan strain of the virus. In this study, we present a new generation of anti-RBD nanobodies with superior properties. The primary representative of this group, Re32D03, neutralizes Alpha to Delta as well as Omicron BA.2.75; other members neutralize, in addition, Omicron BA.1, BA.2, BA.4/5, and XBB.1. Crystal structures of RBD-nanobody complexes reveal how ACE2-binding is blocked and also explain the nanobodies' tolerance to immune escape mutations. Through the cryo-EM structure of the Ma16B06-BA.1 Spike complex, we demonstrated how a single nanobody molecule can neutralize a trimeric spike. We also describe a method for large-scale production of these nanobodies in Pichia pastoris, and for formulating them into aerosols. Exposing hamsters to these aerosols, before or even 24 h after infection with SARS-CoV-2, significantly reduced virus load, weight loss and pathogenicity. These results show the potential of aerosolized nanobodies for prophylaxis and therapy of coronavirus infections.
Keywords: Aerosol; Animal model; COVID-19; Hamster; Inhalation; Nanobodies; Protein structure; Receptor binding domain; SARS-CoV-2; Spike; VHH antibodies; Variants of concern.