RT Journal Article
SR Electronic
T1 Highly potent anti-SARS-CoV-2 multi-DARPin therapeutic candidates
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.08.25.256339
DO 10.1101/2020.08.25.256339
A1 Marcel Walser
A1 Sylvia Rothenberger
A1 Daniel L. Hurdiss
A1 Anja Schlegel
A1 Valérie Calabro
A1 Simon Fontaine
A1 Denis Villemagne
A1 Maria Paladino
A1 Tanja Hospodarsch
A1 Alexandra Neculcea
A1 Andreas Cornelius
A1 Patricia Schildknecht
A1 Mirela Matzner
A1 Martin Hänggi
A1 Marco Franchini
A1 Yvonne Kaufmann
A1 Iris Schlegel
A1 Chloe Iss
A1 Thamar Loser
A1 Susanne Mangold
A1 Christel Herzog
A1 Dieter Schiegg
A1 Christian Reichen
A1 Filip Radom
A1 Andreas Bosshart
A1 Andreas Lehmann
A1 Micha A. Haeuptle
A1 Alexander Zürcher
A1 Toni Vagt
A1 Gabriel Sigrist
A1 Marcel Straumann
A1 Karl Proba
A1 Niina Veitonmäki
A1 Keith M. Dawson
A1 Christof Zitt
A1 Jennifer Mayor
A1 Sarah Ryter
A1 Heyrhyoung Lyoo
A1 Chunyan Wang
A1 Wentao Li
A1 Ieva Drulyte
A1 H. Kaspar Binz
A1 Leon de Waal
A1 Koert J. Stittelaar
A1 Seth Lewis
A1 Daniel Steiner
A1 Frank J.M. van Kuppeveld
A1 Olivier Engler
A1 Berend-Jan Bosch
A1 Michael T. Stumpp
A1 Patrick Amstutz
YR 2020
UL http://biorxiv.org/content/early/2020/08/26/2020.08.25.256339.abstract
AB Globally accessible preventive and therapeutic molecules against SARS-CoV-2 are urgently needed. DARPin molecules are an emerging class of novel therapeutics based on naturally occurring repeat proteins (∼15 kDa in size) and can be rapidly produced in bacteria in large quantities. Here, we report the identification of 380 DARPin molecules specifically targeting the SARS-CoV-2 spike protein selected from a naïve library of 1012 DARPin molecules. Using extensive biophysical and biochemical characterization, (pseudo)virus neutralization assays and cryo-EM analysis, 11 mono-DARPin molecules targeting either the receptor binding domain (RBD), the S1 N-terminal-domain (NTD) or the S2 domain of the SARS-CoV-2 spike protein were chosen. Based on these 11 mono-DARPin molecules, 31 anti-SARS-CoV-2 multi-DARPin molecules were constructed which can broadly be grouped into 2 types; multi-paratopic RBD-neutralizing DARPin molecules and multi-mode DARPin molecules targeting simultaneously RBD, NTD and the S2 domain. Each of these multi-DARPin molecules acts by binding with 3 DARPin modules to the SARS-CoV-2 spike protein, leading to potent inhibition of SARS-CoV-2 infection down to 1 ng/ml (12 pM) and potentially providing protection against viral escape mutations. Additionally, 2 DARPin modules binding serum albumin, conferring an expected half-life of about 3 weeks in humans, were included in the multi-DARPin molecules. The protective efficacy of one multi-DARPin molecule was studied in a Golden Syrian hamster SARS-CoV-2 infection model, resulting in a significant reduction in viral load and pathogenesis. In conclusion, the multi-DARPin molecules reported here display very high antiviral potency, high-production yield, and a long systemic half-life, and thereby have the potential for single-dose use for prevention and treatment of COVID-19.Competing Interest StatementMolecular Partners authors own performance share units and/or stock of the company. HKB owns stock of the company.