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. 2020 Jul 31;11:3859. doi: 10.1038/s41467-020-17698-0

Fig. 2. bCAII and protein responses of supramolecular hydrogels.

Fig. 2

a Schematic illustration of naked eye detection of a gel–sol transition triggered by bCAII and non-enzymatic proteins on a hydrogel array chip. b bCAII response of (left) APmoc-F(CF3)F and (right) Bz-FF. EZA: ethoxzolamide. Condition for bCAII response: [APmoc-F(CF3)F] = 0.35 wt% (6.1 mM), [Bz-FF] = 2.0 wt% (42 mM), [bCAII] = 10 µM, [EZA] = 100 µM, 100 mM HEPES (pH 8.0), 25 °C, 6 h, Vgel:Vstimulus = 10:1. c Avidin response of APmoc-F(CF3)F hydrogels containing bCAII and EAT(avidin). d Residual ratios of APmoc-F(CF3)F after treatment of avidin determined by HPLC. The background hydrolysis of the acetyl group occurred probably because the enzymatic activity of bCAII was not completely inhibited (Supplementary Fig. 7). e High-resolution Airyscan CLSM images of APmoc-F(CF3)F hydrogels containing bCAII and EAT(avidin) with a fluorescent probe, TMR-Gua, after addition of (left) buffer, (middle) avidin, and (right) the mixture of avidin and biotin. f Determination of the detection threshold. g The non-enzymatic protein response in various combinations of target proteins and EATs. APmoc-F(CF3)F hydrogels changed to the sol state only under the appropriate pairs. h Heat map of the residual ratios of APmoc-F(CF3)F after addition of analytes determined by HPLC analysis. CGC: Critical gelation concentration. Condition for non-enzymatic protein response: [APmoc-F(CF3)F] = 0.35 wt% (6.1 mM), [bCAII] = 10 µM, [EAT(avidin)] = 20 µM, [EAT(DHFR)] = 45 µM, [EAT(DNP-IgG)] = 15 µM, [avidin] = 20 µM (for c, d, e, g, and h) and 0, 1.25, 2.5, 5.0, 10, 20 µM (for f), [biotin] = 120 µM, [DHFR] = 45 µM (for g, h), [DNP IgG] = 15 µM (for g, h), [TMR-Gua] = 10 µM (for e), 100 mM HEPES, pH 8.0, 25 °C, 6 (for c, d, and e), 12 (for f), and 18 h (for g and h), Vgel:Vstimulus = 10:1.