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. Author manuscript; available in PMC: 2024 Jan 22.
Published in final edited form as: Cell Rep. 2023 Dec 13;42(12):113553. doi: 10.1016/j.celrep.2023.113553

Figure 1. Design and immunogenicity of hyperglycosylated NC99 trihead nanoparticle immunogens.

Figure 1.

(A) Model structures and gene diagrams for wild-type and hyperglycosylated NC99 triheads with wild-type glycans in light purple and glycan knockins in dark purple. NC99 HA numbering is in blue, and trihead model numbering is in black.

(B) Reducing SDS-PAGE of wild-type and hyperglycosylated NC99 monoheads and triheads without and with PNGaseF digestion.

(C) BLI of wild-type and hyperglycosylated NC99 monoheads and triheads against C05, FluA-20, and Ab6649.

(D) nsEM micrographs of hyperglycosylated NC99 monohead and trihead I53_dn5 nanoparticles. Scale bars = 100 nm.

(E) Schematic illustrating mouse study timeline, immunizations, and serology timepoint.

(F) Week 10 NC99-foldon trimer ELISA titers plotted as the reciprocal EC50 titer, hemagglutination inhibition (HAI) titers, and the ratio of HAI/reciprocal EC50 titers of hyperglycosylated NC99 monohead and trihead nanoparticles in BALB/c mice. Each symbol represents an individual animal, and the geometric mean of each group is indicated by the bar (n = 5 mice/group).

Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test; *p < 0.05; **p < 0.01.