1. Nanobodies to multiple spike variants and inhalation of nanobody-containing aerosols neutralize SARS-CoV-2 in cell culture and hamsters.
- Author
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Aksu, Metin, Kumar, Priya, Güttler, Thomas, Taxer, Waltraud, Gregor, Kathrin, Mußil, Bianka, Rymarenko, Oleh, Stegmann, Kim M., Dickmanns, Antje, Gerber, Sabrina, Reineking, Wencke, Schulz, Claudia, Henneck, Timo, Mohamed, Ahmed, Pohlmann, Gerhard, Ramazanoglu, Mehmet, Mese, Kemal, Groß, Uwe, Ben-Yedidia, Tamar, and Ovadia, Oded
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MICROBIOLOGICAL aerosols , *HAMSTERS , *CORONAVIRUS diseases , *COVID-19 , *SARS-CoV-2 , *CELL culture , *CHO cell , *IMMUNOGLOBULINS - Abstract
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. [Display omitted] • Nanobodies to Spike proteins neutralize recent SARS-CoV-2 variants. • Nanobody-spike structures explain mutation tolerance. • Vaporizing technology allows nanobody application as aerosol. • Prophylactic inhalation suppresses infection below detectability. • Nanobodies diminish weight loss even when applied 24 h after infection. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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