Figure 6.

ConSOSL.UFO.664 Env Trimers Present a Native-Like Pre-fusion Conformation

(A) Size exclusion chromatography (SEC) profile of the ConSOSL.UFO.664 produced in 293T.17 cells.

(B) Native PAGE-western blotting of SEC fractions from (A), detected with 2G12.

(C) Hydrophilic interaction-ultra-performance liquid chromatography (HILIC-UPLC) profile of N-linked glycans from SEC purified ConSOSL.UFO.664 trimers. Oligomannose-type (green) and complex-type (pink) glycans were identified by endoglycosidase H (Endo H) digestion. Rel., relative.

(D) Relative N-glycosylation sites.

(E) mAbs titration against ConSOSL.UFO.664 Myc-HIS trimers captured by anti-cMyc 9E10 Ab. Representative of at least n = 2 independent experiments. The titration curves are grouped by Env domain specificity, and 2G12 is plotted on all of the graphs for comparison. Solid lines, bNAbs; dotted lines, nNAbs. OD, optical density.

(F) Same as in (E) for gp41-specific mAbs.

(G and H) Negative stain-electron microscopy (NS-EM) images showing views of (G) the propeller shape of the ligand-free trimer and (H) in complex with VRC01 Fab.

See also Figures S6B–S6I.