Production and purification of recombinant SARS COV-2 RBDSD1-Fc protein from a transiently transfected EXPI293 mammalian cells.
Expression plasmid for CoV-9-RBDSD1, residues of RBD R319−S591 with a C-terminal HRV3C protease cleavage site, a monomeric Fc tag and an 8XHisTag was generously gifted by Dr. McLellan’s lab (1). Secreted proteins were purified from a suspension culture cell supernatant with an expression level 15-20mg/L. For different application RBD-SD1 was purified by affinity and gel-filtration protocols, with and without purification tag (Figure 1.)
Reference: 1.Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Daniel Wrapp, Nianshuang Wang, Kizzmekia S. Corbett,A. Goldsmith, Ching-Lin Hsieh,Olubukola Abiona, Barney S. Graham, Jason S. McLellan .Science 367, 1260–1263 (2020) 13 March 2020