Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications
Authors: Altincekic Nadide , Korn Sophie Marianne , Qureshi Nusrat Shahin , Dujardin Marie , Ninot-Pedrosa Martí , Abele Rupert , Abi Saad Marie Jose , Alfano Caterina , Almeida Fabio C. L. , Alshamleh Islam , de Amorim Gisele Cardoso , Anderson Thomas K. , Anobom Cristiane D. , Anorma Chelsea , Bains Jasleen Kaur , Bax Adriaan , Blackledge Martin , Blechar Julius , Böckmann Anja , Brigandat Louis , Bula Anna , Bütikofer Matthias , Camacho-Zarco Aldo R. , Carlomagno Teresa , Caruso Icaro Putinhon , Ceylan Betül , Chaikuad Apirat , Chu Feixia , Cole Laura , Crosby Marquise G. , de Jesus Vanessa , Dhamotharan Karthikeyan , Felli Isabella C. , Ferner Jan , Fleischmann Yanick , Fogeron Marie-Laure , Fourkiotis Nikolaos K. , Fuks Christin , Fürtig Boris , Gallo Angelo , Gande Santosh L. , Gerez Juan Atilio , Ghosh Dhiman , Gomes-Neto Francisco , Gorbatyuk Oksana , Guseva Serafima , Hacker Carolin , Häfner Sabine , Hao Bing , Hargittay Bruno , Henzler-Wildman K. , Hoch Jeffrey C. , Hohmann Katharina F. , Hutchison Marie T. , Jaudzems Kristaps , Jović Katarina , Kaderli Janina , Kalniņš Gints , Kaņepe Iveta , Kirchdoerfer Robert N. , Kirkpatrick John , Knapp Stefan , Krishnathas Robin , Kutz Felicitas , zur Lage Susanne , Lambertz Roderick , Lang Andras , Laurents Douglas , Lecoq Lauriane , Linhard Verena , Löhr Frank , Malki Anas , Bessa Luiza Mamigonian , Martin Rachel W. , Matzel Tobias , Maurin Damien , McNutt Seth W. , Mebus-Antunes Nathane Cunha , Meier Beat H. , Meiser Nathalie , Mompeán Miguel , Monaca Elisa , Montserret Roland , Mariño Perez Laura , Moser Celine , Muhle-Goll Claudia , Neves-Martins Thais Cristtina , Ni Xiamonin , Norton-Baker Brenna , Pierattelli Roberta , Pontoriero Letizia , Pustovalova Yulia , Ohlenschläger Oliver , Orts Julien , Da Poian Andrea T. , Pyper Dennis J. , Richter Christian , Riek Roland , Rienstra Chad M. , Robertson Angus , Pinheiro Anderson S. , Sabbatella Raffaele , Salvi Nicola , Saxena Krishna , Schulte Linda , Schiavina Marco , Schwalbe Harald , Silber Mara , Almeida Marcius da Silva , Sprague-Piercy Marc A. , Spyroulias Georgios A. , Sreeramulu Sridhar , Tants Jan-Niklas , Tārs Kaspars , Torres Felix , Töws Sabrina , Treviño Miguel Á. , Trucks Sven , Tsika Aikaterini C. , Varga Krisztina , Wang Ying , Weber Marco E. , Weigand Julia E. , Wiedemann Christoph , Wirmer-Bartoschek Julia , Wirtz Martin Maria Alexandra , Zehnder Johannes , Hengesbach Martin , Schlundt Andreas 
Published: May 2021

The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.