作者
Natalia G Herrera, Nicholas C Morano, Alev Celikgil, George I Georgiev, Ryan J Malonis, James H Lee, Karen Tong, Olivia Vergnolle, Aldo B Massimi, Laura Y Yen, Alex J Noble, Mykhailo Kopylov, Jeffrey B Bonanno, Sarah C Garrett-Thomson, David B Hayes, Robert H Bortz III, Ariel S Wirchnianski, Catalina Florez, Ethan Laudermilch, Denise Haslwanter, J Maximilian Fels, M Eugenia Dieterle, Rohit K Jangra, Jason Barnhill, Amanda Mengotto, Duncan Kimmel, Johanna P Daily, Liise-anne Pirofski, Kartik Chandran, Michael Brenowitz, Scott J Garforth, Edward T Eng, Jonathan R Lai, Steven C Almo
发表日期
2020/12/21
期刊
ACS omega
卷号
6
期号
1
页码范围
85-102
出版商
American Chemical Society
简介
Coronavirus disease 2019 (COVID-19) is a global health crisis caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and there is a critical need to produce large quantities of high-quality SARS-CoV-2 Spike (S) protein for use in both clinical and basic science settings. To address this need, we have evaluated the expression and purification of two previously reported S protein constructs in Expi293F and ExpiCHO-S cells, two different cell lines selected for increased protein expression. We show that ExpiCHO-S cells produce enhanced yields of both SARS-CoV-2 S proteins. Biochemical, biophysical, and structural (cryo-EM) characterizations of the SARS-CoV-2 S proteins produced in both cell lines demonstrate that the reported purification strategy yields high-quality S protein (nonaggregated, uniform material with appropriate biochemical and biophysical properties), and analysis of …
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NG Herrera, NC Morano, A Celikgil, GI Georgiev… - ACS omega, 2020