An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike
Michael Schoof,Bryan Faust,R.A. Saunders,Smriti Sangwan,Veronica V. Rezelj,Nick Hoppe,Morgane Boone,Christian B. Billesbølle,Cristina Puchades,Caleigh M. Azumaya,Huong T. Kratochvil,M. Zimanyi,Ishan Deshpande,Jiahao Liang,S. Dickinson,Henry C. Nguyen,Cynthia M. Chio,Gregory E. Merz,Michael C. Thompson,Devan Diwanji,Kaitlin Schaefer,Aditya A. Anand,Niv Dobzinski,Beth S. Zha,Camille R. Simoneau,Camille R. Simoneau,Kristoffer E. Leon,Kristoffer E. Leon,Kris M. White,Un Seng Chio,Meghna Gupta,Mingliang Jin,Fei Li,Yanxin Liu,Kaihua Zhang,David Bulkley,Ming Sun,Amber M. Smith,Alexandrea N. Rizo,Frank R. Moss,Axel F. Brilot,Sergei Pourmal,Raphael Trenker,Thomas H. Pospiech,Sayan Gupta,Benjamin Barsi-Rhyne,Vladislav Belyy,A.W. Barile-Hill,Silke Nock,Yuwei Liu,Nevan J. Krogan,Corie Y. Ralston,Danielle L. Swaney,Adolfo García-Sastre,Melanie Ott,Melanie Ott,Marco Vignuzzi,Peter Walter,Aashish Manglik +58 more
TLDR
Nanobodies that bind tightly to spike and efficiently neutralize SARS-CoV-2 in cells are reported, which enables aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia.Abstract:
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus enters host cells via an interaction between its Spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2). By screening a yeast surface-displayed library of synthetic nanobody sequences, we developed nanobodies that disrupt the interaction between Spike and ACE2. Cryo-electron microscopy (cryo-EM) revealed that one nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains locked into their inaccessible down state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains function after aerosolization, lyophilization, and heat treatment, which enables aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia.read more
Citations
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Journal ArticleDOI
Generation of Photocaged Nanobodies for Intracellular Applications in an Animal Using Genetic Code Expansion and Computationally Guided Protein Engineering
Jack M O'Shea,Angeliki Goutou,Jack Brydon,Cyrus R. Sethna,Christopher W. Wood,Sebastian Greiss +5 more
TL;DR: This work develops a general approach to engineer photo‐activatable nanobodies using photocaged amino acids that are introduced into the target binding interface by genetic code expansion, and tunes nanobody/target binding affinity to eliminate binding before uncaging.
Posted Content
Nanobodies recognizing conserved hidden clefts of all SARS-CoV-2 spike variants
Ryota Maeda,Junso Fujita,Yoshinobu Konishi,Yasuhiro Kazuma,Hiroyuki Yamazaki,Itsuki Anzai,Keishi Yamaguchi,Kazuki Kasai,Kayoko Nagata,Yutaro Yamaoka,Kei Miyakawa,Akihide Ryo,Kotaro Shirakawa,Fumiaki Makino,Yoshiharu Matsuura,Tsuyoshi Inoue,Akihiro Imura,Keiichi Namba,Akifumi Takaori-Kondo +18 more
TL;DR: In this paper, the sequences of nine alpaca nanobodies were used to detect the spike proteins of four SARS-CoV-2 variants of concern (VOCs), namely alpha, beta, gamma, and delta variants.
Journal ArticleDOI
Localized delivery of nanomedicine and antibodies for combating COVID-19
TL;DR: In this paper , a review of the inhaled nanomedicines and antibodies, as well as intranasal nanodrugs, for the prevention and treatment of COVID-19 are summarized.
Book ChapterDOI
Exploring the use of intracellular and extracellular allosteric modulators to understand GPCR signaling
Kyla Bourque,Juliana C.C. Dallagnol,Hassan Nassour,David Chatenet,Bruce G. Allen,Terence E. Hébert +5 more
TL;DR: In this article , the authors discuss how intracellular allosteric modulators, such as intercellular ions, peptidomimetic ligands or pepducins generated from the primary and secondary structures of GPCRs, have also been used as key tools to understand and drive GPCR signaling.
Journal ArticleDOI
Expanding and improving nanobody repertoires using a yeast display method: Targeting SARS-CoV-2
Frederick R. Cross,Peter C. Fridy,Natalia E. Ketaren,Fred D. Mast,S. Li,Pernet Olivier,Kresti Pecani,Brian T. Chait,John D. Aitchison,Michael P. Rout +9 more
TL;DR: Yeast display has been used to identify and characterize a broad spectrum of anti-SARS-CoV-2 Spike nanobodies using B-cell complementary DNA from immunized animals as a source of VHH sequences as mentioned in this paper .
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