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. 2020 Jun 5;17(7):2734–2748. doi: 10.1021/acs.molpharmaceut.0c00419

Figure 3.

Figure 3

Biophysical analysis of FGF2 conjugates. (A) Fluorescence emission spectra (300–450 nm) of FGF2 conjugates at a concentration of 4 μM upon excitation at 280 nm. The dashed line represents FGF2 unfolded in 6 M GdmCl. (B) Thermal denaturation of FGF2 conjugates at a concentration of 3 μM, monitored by the change in the fluorescence at 350 nm. (C) Analysis of the aggregation of FGF2 conjugates evaluated by changes in light back-reflection during thermal denaturation. (D) Retention volume of FGF2 conjugates determined by size-exclusion chromatography. Estimated molecular weights are given in Table 1. (E) Stability of FGF2 and FGF2 conjugates in human serum. The FGF2 and the conjugates were incubated at a concentration of 1 μg/mL without heparin in the human serum at 37 °C for indicated times. The samples were analyzed by SDS-PAGE and immunoblotting using the anti-FGF2 antibody. Experiments were performed in triplicate, and representative results are shown.