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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Mol Pharm. 2019 May 30;16(7):3024–3039. doi: 10.1021/acs.molpharmaceut.9b00263

Figure 2.

Figure 2.

High molecular weight FAF–Rapa has the purity, size, and concentration–temperature phase behavior necessary for stability at body temperature. (A) Identity, purity, and fluorescence of FAF, FAF–Rapa, and rhodamine-labeled FAF–Rapa (Rho–FAF–Rapa) were analyzed by Coomassie blue staining and fluorescence imaging of SDS–PAGE. (B) Dynamic light scattering shows that all three formulations (Table 1) remain monodisperse at 37 °C (10 μM, PBS). (C) Optical density of FAF was monitored as a function of temperature at 350 nm to confirm solubility in PBS at physiological temperatures (shaded area). The Tt of FAF at 25 μM was 57.0 °C. (D) Using optical density, the phase transition temperature was plotted vs. concentration as a phase diagram, below which FAF remains soluble (n = 3, Mean ± SD). The shaded area indicates physiological temperatures. Dotted lines show a 95% confidence interval (CI) of the mean.