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. 2023 Jan 21;24(3):2161. doi: 10.3390/ijms24032161

Figure 9.

Figure 9

MST measurements of TF2D-5, TF2D-5a, and CP1 with full-length tau. (A) Boltzmann fit of TF2D-5 MST measurements with full-length tau. TF2D-5 was labeled with CF633. The concentration of TF2D-5 was 500 nM, while the concentration of full-length tau varied between 2.5 µM and 0.2 pM. The measurement was performed in 1× PBS pH 7.4 containing 150 mM NaCl. (B) Boltzmann fit of TF2D-5a MST measurements with full-length tau. TF2D-5a was labeled with CF633. The concentration of TF2D-5a was 62.5 nM, while the concentration of full-length tau varied between 300 nM and 0.5 nM. The measurement was performed in 0.1 M sodium phosphate buffer pH 7.4 containing 75 mM sodium sulfate and 0.05% Tween20. (C) Average of CP1 MST measurements with full-length tau. CP1 was labeled with CF633. The concentration of CP1 was 62.5 nM, while the concentration of full-length tau varied between 2.5 µM and 0.2 pM. The measurement was performed in 0.1 M sodium phosphate buffer pH 7.4 containing 75 mM sodium sulfate and 0.05% Tween 20. (D) Calculated KDs of each peptide and full-length tau. TF2D-5 has a KD of 308 pM with an error of 148 pM to full-length tau, which is tenfold lower that the KD determined by SPR measurements. TF2D-5a has a KD of 11.7 nM with an error of 1.3 nM. This is close to the KD determined by SPR measurements (6.1 nM). For CP1, no fit could be performed, and all measurements indicated no binding in the tested concentration range. All experiments were repeated four times and the average of all measurements was calculated, which in turn was used to calculate the fit (Boltzmann).