FIG 10.

Glycan composition of SOSIP.664-His and gp140_UNC-Fd-His proteins. HILIC-UPLC chromatograms of fluorescently labeled N-linked glycans released from 92UG037.8 and CZA97.012 SOSIP.664-His and gp140_UNC-Fd-His proteins, respectively, purified by the bNAb/SEC and Ni-NTA/SEC methods. The Foldon sequence present in the gp140_UNC-Fd-His proteins does not contain any canonical NXT or NXS motifs, so it is highly unlikely to be glycosylated. Glycans were released from reducing SDS-polyacrylamide gel bands using PNGase F. Peaks corresponding to oligomannose glycans (Man_5–9GlcNAc₂) are colored in shades of green; the remaining peaks (gray) correspond to complex and hybrid-type glycans. Peak areas (as a percentage of total glycans) are recorded as numerical values in the associated pie charts and should be consulted to obtain a quantitative estimate of the differences in the glycan contents of the comparator proteins. Glycan structures are depicted according to the color scheme established by the Consortium for Functional Genomics (http://www.functionalglycomics.org/): green circles for mannose and blue squares for N-acetylglucosamine.