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. 2017 Dec;106(12):3474–3485. doi: 10.1016/j.xphs.2017.07.019

Figure 3.

Figure 3

HOS and overall conformational stability of dmLT in formulation buffer (see Methods and Materials section) at pH 7.4. (a) Representative second derivative FTIR spectrum of dmLT in the amide I region (see Supplemental Table S1 for individual peak assignments including SD values, n = 3), (b) far-UV CD spectrum at 10°C, (c) thermal melt for intrinsic Trp fluorescence peak position (10°C to 90°C); the inset shows intrinsic tryptophan fluorescence spectra's at 10°C and 90°C, (d) static light scattering at 295 nm (10°C to 90°C), (e) extrinsic ANS fluorescence peak intensity (10°C to 90°C); the inset shows the extrinsic ANS fluorescence spectra at 10°C and 90°C. Two major transitions as indicated by arrows, 1 for A chain (∼50°C) and the second for B chain (∼70°C) are observed using a variety of biophysical measurements. (f) Representative DSC thermogram of dmLT. The black line represents raw thermogram, and the red line represents the curve fitted data to 4 peaks. The Tm1, Tm2 values indicate the thermal melting transitions for the A chain and Tm3, Tm4 values indicate the thermal melting transitions for the B chain of dmLT. Error bars are SD from triplicate experiments.