Structure-guided multivalent nanobodies block SARS-CoV-2 infection and suppress mutational escape

Jonathan L. Schmid‐Burgk, Ian A. Wilson, Hiroki Kato, Paul-Albert Koenig, Beate M. Kümmerer, Maria H. Christensen, Yonas M. Tesfamariam, Florian I. Schmidt, Kerstin U. Ludwig, Nicholas C. Wu, Matthias Geyer, Meng Yuan, Karl Gatterdam, Michael Schindler, Xueyong Zhu, Natalia Ruétalo, Hrishikesh Das, Hejun Liu, Florian N. Gohr, Lea‐Marie Jenster, Lisa D. J. Schiffelers, Miki Uchima, Jennifer Deborah Wuerth, Caroline I. Fandrey, Sabine Normann, Jan M. P. Tödtmann, Steffen Pritzl, Leo Hanke, Jannik Boos, B.M. Hallberg
2021-01-12

SCID:  54.1/yx22xexf
The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread, with devastating consequences. For passive immunization efforts, nanobodies have size and cost advantages over conventional antibodies. In this study, we generated four neutralizing nanobodies that target the receptor binding domain of the SARS-CoV-2 spike protein. We used x-ray crystallography and cryo-electron microscopy to define two distinct binding epitopes. On the basis of these structures, we engineered multivalent nanobodies with more than 100 times the neutralizing activity of monovalent nanobodies. Biparatopic nanobody fusions suppressed the emergence of escape mutants. Several nanobody constructs neutralized through receptor binding competition, whereas other monovalent and biparatopic nanobodies triggered aberrant activation of the spike fusion machinery. These premature conformational changes in the spike protein forestalled productive fusion and rendered the virions noninfectious.
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2021-01-12
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Authors
Jonathan L. Schmid‐Burgk
Ian A. Wilson
Hiroki Kato
Paul-Albert Koenig
Beate M. Kümmerer
Maria H. Christensen
Yonas M. Tesfamariam
Florian I. Schmidt
Kerstin U. Ludwig
Nicholas C. Wu
Matthias Geyer
Meng Yuan
Karl Gatterdam
Michael Schindler
Xueyong Zhu
Natalia Ruétalo
Hrishikesh Das
Hejun Liu
Florian N. Gohr
Lea‐Marie Jenster
Lisa D. J. Schiffelers
Miki Uchima
Jennifer Deborah Wuerth
Caroline I. Fandrey
Sabine Normann
Jan M. P. Tödtmann
Steffen Pritzl
Leo Hanke
Jannik Boos
B.M. Hallberg
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