Fig. 3: Glycan analysis of BG505 Env immunogens with wildtype and modified glycans. | Nature Communications

Fig. 3: Glycan analysis of BG505 Env immunogens with wildtype and modified glycans.

From: Single-component multilayered self-assembling protein nanoparticles presenting glycan-trimmed uncleaved prefusion optimized envelope trimers as HIV-1 vaccine candidates

Fig. 3: Glycan analysis of BG505 Env immunogens with wildtype and modified glycans.The alternative text for this image may have been generated using AI.

a Comparison of site-specific occupancy and oligomannose-type glycan content for the BG505 UFO trimer (wildtype, swainsonine, and kifunensine) and three SApNPs (wildtype). The plotted data represent the mean of three analytical repeats, with trimers shown as circles and NPs as squares. The oligomannose content was determined by adding all glycan compositions containing HexNAc(2). b SEC profiles of BG505 UFO-E2p-L4P and UFO-10GS-I3-01v9-L7P SApNPs upon endo H treatment. c Reducing SDS-PAGE of BG505 UFO-E2p-L4P and UFO-10GS-I3-01v9-L7P SApNPs before and after glycan trimming by endo H. d EM images of glycan-trimmed BG505 UFO-E2p-L4P and UFO-10GS-I3-01v9-L7P SApNPs. e Thermostability of glycan-trimmed BG505 UFO-E2p-L4P and UFO-10GS-I3-01v9-L7P SApNPs, with Tm, ΔT1/2, and Ton measured by DSC. f Reducing SDS-PAGE of BG505 UFO-E2p-L4P and UFO-10GS-I3-01v9-L7P SApNPs before and after endo H treatment under two different conditions: 25 °C for 5 h (condition 1) and protein denaturation at 100 °C followed by treatment at 37 °C for 1 h (condition 2). g UPLC analysis of released glycans from BG505 UFO-10GS-I3-01v9 SApNP. Top: natively glycosylated; Middle: endo H-treated; Bottom: overlay of two UPLC traces with the treated trace (from the middle panel) recolored in white. The endo H-treated trace has been normalized to the untreated trace using the complex-type glycan peak eluting at ~8 min. In top and middle panels: oligomannose-type and complex-type glycans are colored in green and pink, respectively. In the top panel, oligomannose-type glycan peaks are labeled with their representative structures. For both UPLC traces, the percentage of each oligomannose-type is listed on the right.

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