Recombinant gp140 Envs were expressed as chimeric SOSIPs, where the gp120 of CH0848 was grafted onto the BG505 gp41 [88 (link)]. The trimers were stabilized with E64K and A316W mutations [15 (link), 49 (link)]. Proteins were expressed in Freestyle293 cells (Invitrogen) using transient cotransfection of Env and furin DNA. Cell-free culture media was subjected to PGT151 antibody affinity and superdex200 size exclusion chromatography.
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