Fig. 5
Glycosylation on the uncleaved, soluble NFL Env trimer. a On the left, superimposition of oligomannose glycans (green) observed on the BG505 NFL.664 trimer (shades of gray) versus the non-deglycosylated BG505 SOSIP.664 (red; PDB 5FYL) illustrates the protection by the bound antibodies from EndoH trimming, and the glycan fence around the PGV19 footprint (LCvar: cyan, HCvar: orange). On the right are various conformations of well-defined glycan sub- or super-structures observed in the BG505 NFL.664 crystal structure. b HILIC-UPLC spectra of fluorescently labeled N-linked glycans released from BG505 NFL.664 and BG505 SOSIP.664 produced in HEK 293F cells. Oligomannose and hybrid glycans are colored in green and complex glycans in pink. The corresponding structures for the oligomannose glycans (Man5-9GlcNAc2) are shown in the upper panel. c Site-specific N-glycosylation analysis of BG505 NFL.664. Relative quantification of the micro-heterogeneity of 20 out of the 27 N-glycosylation sites of BG505 NFL.664 produced in HEK 293F cells were determined, with the color scheme preserved from panel b. Glycoform annotation: oligomannose glycans Man5GlcNAc2 to Man9GlcNAc2 are represented as M5 to M9, hybrids as H, and fucosylated hybrids as FH. Complex glycans are also categorized by the number of branching antennae (An); number of galactose residues (Gn); and core fucose (f)
