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. Author manuscript; available in PMC: 2019 Jan 9.
Published in final edited form as: Nat Biotechnol. 2018 Jul 9;36(8):707–716. doi: 10.1038/nbt.4181

Figure 1. Synthesis and characterization of TCR signalling-responsive protein nanogels.

Figure 1

(a) Naïve or con-A-primed CD8+ T cells were incubated in the presence of gp100 peptide (10 μg/mL) or anti-CD3/CD28 beads for 24 hrs followed by measurement of WST-1 cell-surface reduction rate in presence of an intermediate electron acceptor for 1 hr at 37°C. (b) Con-A-primed CD8+ T cells were incubated in the presence of anti-CD3/CD28 beads and the cell-surface reduction rate was measured over time. (c) Proposed strategy for linking elevated surface redox activity of activated CD8+ T cells to accelerated drug release kinetics from a redox-responsive backpack. (d) Scheme for protein nanogel (NG) synthesis, and release of protein in response to reducing activity in the local microenvironment. (e) Representative TEM image of NGs prepared from IL-15Sa. (f) Mean ± s.d. hydrodynamic sizes of different NGs determined by dynamic light scattering (n=3 independent samples). (g) Release kinetics of cytokines from redox-responsive or non-degradable IL-15Sa-NGs in PBS with or without added glutathione (GSH) as a reducing agent. (h) Released and native cytokines were characterized by MALDI mass spectrometry. Data in a, b represent the mean ± s.e.m. (n = 3 biologically independent samples /group) and analysed by One-Way ANOVA and Tukey’s tests. All data are one representative of at least two independent experiments.