Nanobodies to multiple spike variants and inhalation of nanobody-containing aerosols neutralize SARS-CoV-2 in cell culture and hamsters

被引：13

作者：

Aksu, Metin

Kumar, Priya ^{[1
]}

Guettler, Thomas ^{[2
]}

Taxer, Waltraud

Gregor, Kathrin

Mussil, Bianka ^{[1
]}

Rymarenko, Oleh

Stegmann, Kim M. ^{[1
]}

Dickmanns, Antje ^{[1
]}

Gerber, Sabrina ^{[1
]}

Reineking, Wencke ^{[3
]}

Schulz, Claudia ^{[4
]}

Henneck, Timo ^{[4
,5
]}

Mohamed, Ahmed ^{[4
,5
]}

Pohlmann, Gerhard ^{[6
]}

Ramazanoglu, Mehmet ^{[6
]}

Mese, Kemal ^{[7
]}

Gross, Uwe ^{[8
]}

Ben-Yedidia, Tamar ^{[9
]}

Ovadia, Oded ^{[9
]}

Fischeri, Dalit Weinstein ^{[9
]}

Kamensky, Merav ^{[9
]}

Reichman, Amir ^{[9
]}

Baumgartner, Wolfgang ^{[4
]}

von Koeckritz-Blickwede, Maren ^{[5
,6
]}

Dobbelstein, Matthias ^{[1
,2
]}

Goerlich, Dirk ^{[1
]}

机构：

[1] Max Planck Inst Multidisciplinary Sci, Dept Cellular Logist, Fassberg 11, D-37077 Gottingen, Germany

[2] Univ Med Ctr Gottingen, Dept Mol Oncol, Inst Mol Oncol, Justus von Liebig Weg 11, D-37077 Gottingen, Germany

[3] Octapharma Biopharmaceut GmbH, Neuenheimer Feld 590, D-69220 Heidelberg, Germany

[4] Univ Vet Med Hannover, Dept Pathol, Bunteweg 17, D-30559 Hannover, Germany

[5] Univ Vet Med Hannover, Res Ctr Emerging Infect & Zoonosis RIZ, Bunteweg 17, D-30559 Hannover, Germany

[6] Univ Vet Med Hannover, Dept Biochem, Bunteweg 17, D-30559 Hannover, Germany

[7] Fraunhofer Inst Toxicol & Expt Med ITEM, Nikolai Fuchs Str 1, D-30625 Hannover, Germany

[8] Univ Med Ctr Gottingen, Dept Med Microbiol & Virol, Kreuzbergring 57, D-37075 Gottingen, Germany

[9] Scinai Immunotherapeut Ltd, Jerusalem Biopk, IL-9112001 Jerusalem, Israel

来源：

ANTIVIRAL RESEARCH | 2024年 / 221卷

关键词：

SARS-CoV-2; Variants of concern; Spike; Receptor binding domain; Nanobodies; VHH antibodies; Protein structure; Inhalation; Aerosol; COVID-19; Animal model; Hamster; STRUCTURE REFINEMENT; PROTEIN; SYSTEM; EFFICIENT; SELECTION; RECEPTOR; POTENT;

D O I：

10.1016/j.antiviral.2023.105778

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

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.

引用

页数：21

共 80 条

[11] Structural and functional characterizations of infectivity and immune evasion of SARS-CoV-2 Omicron [J].