Figure 2. AFRs mediate target‐specific delivery of TNF activity.

• A
  Schematic representation of AFR, consisting of N‐terminal VHH fused via a 20xGGS linker to an engineered TNF (e‐TNF) variant and C‐terminal affinity tag.
• B, C
  Viability of L929 parental or mCD20‐expressing cells after 72‐h stimulation with mTNF or sc mTNF mutant Y86F fused to a BcII10 or mCD20 VHH (B) and viability of MCF7 parental or hCD20‐expressing cells after 72‐h stimulation with hTNF or sc hTNF mutant Y87F fused to a BcII10 or hCD20 VHH (C). Cell viability was measured via an ATP luminescence assay. Each point is the mean of three replicates, and error bars are SEM.
• D
  IL‐8 concentration in HUVEC supernatant after 24‐h stimulation with the indicated concentration of stimulus, measured via ELISA. Error bars are SEM. ns, non‐significant; **P < 0.01; ***P < 0.001 by one‐way ANOVA with Bonferroni's multiple comparison test.

Source data are available online for this figure.