Issue link: https://resources.nanotempertech.com/i/1050515
7 APPLICATION NOTE ©2017 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved. 100 150 200 250 300 Scattering (mAU) 80 100 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90 100 0.76 0.78 0.80 0.82 0.84 0.86 0.88 0.90 First Derivative 0 60 80 100 F01 F02 F03 F04 F05 F350/F330 0 60 70 80 90 Temperature (°C) Temperature (°C) Tagg (°C) Agg at 95°C (mAU) T m 1 onset (°C) T m 1 (°C) T m 2 (°C) F01 81.19 +/- 0.05 313.46 59.46 +/- 0. 25 69.69 +/- 0.02 80.36 +/- 0.22 F02 80.82 +/- 0.03 312.06 59.31 +/- 0.10 69.74 +/- 0.07 80.83 +/- 0.22 F03 85.96 +/- 0.07 279.32 66.06 +/- 0.18 75.33 +/- 0.09 84.28 +/- 0.06 F04 86.41 +/- 0.02 225.24 64.24 +/- 0.01 73.82 +/- 0.05 84.80 +/- 0.55 F05 82.77 +/- 0.00 243.26 55.98 +/- 0.11 66.86 +/- 0.08 81.13 +/- 0.15 F01 F02 F03 F04 F05 Thermal unfolding Unfolding-induced aggregation 100 150 200 250 300 Scattering (mAU) 80 100 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90 100 0.76 0.78 0.80 0.82 0.84 0.86 0.88 0.90 First Derivative 0 60 80 100 F01 F02 F03 F04 F05 F350/F330 0 60 70 80 90 Temperature (°C) Temperature (°C) Tagg (°C) Agg at 95°C (mAU) T m 1 onset (°C) T m 1 (°C) T m 2 (°C) F01 81.19 +/- 0.05 313.46 59.46 +/- 0. 25 69.69 +/- 0.02 80.36 +/- 0.22 F02 80.82 +/- 0.03 312.06 59.31 +/- 0.10 69.74 +/- 0.07 80.83 +/- 0.22 F03 85.96 +/- 0.07 279.32 66.06 +/- 0.18 75.33 +/- 0.09 84.28 +/- 0.06 F04 86.41 +/- 0.02 225.24 64.24 +/- 0.01 73.82 +/- 0.05 84.80 +/- 0.55 F05 82.77 +/- 0.00 243.26 55.98 +/- 0.11 66.86 +/- 0.08 81.13 +/- 0.15 F01 F02 F03 F04 F05 Thermal unfolding Unfolding-induced aggregation Figure 4: Analysis of mAb stability by nanoDSF. Thermal unfolding curves of mAb1 in formulations F01-F05. Changes in the F350/F330 fluorescence ratio (top) and the corresponding first derivative are shown. T m - and T m onset values represent averages with standard deviation from triplicate measurements. 100 150 200 250 300 Scattering (mAU) 80 100 20 30 40 50 60 70 80 90 0 10 20 30 40 50 60 70 80 90 100 0.76 0.78 0.80 0.82 0.84 0.86 0.88 0.90 First Derivative 0 60 80 100 F01 F02 F03 F04 F05 F350/F330 0 60 70 80 90 Temperature (°C) Temperature (°C) Tagg (°C) Agg at 95°C (mAU) T m 1 onset (°C) T m 1 (°C) T m 2 (°C) F01 81.19 +/- 0.05 313.46 59.46 +/- 0. 25 69.69 +/- 0.02 80.36 +/- 0.22 F02 80.82 +/- 0.03 312.06 59.31 +/- 0.10 69.74 +/- 0.07 80.83 +/- 0.22 F03 85.96 +/- 0.07 279.32 66.06 +/- 0.18 75.33 +/- 0.09 84.28 +/- 0.06 F04 86.41 +/- 0.02 225.24 64.24 +/- 0.01 73.82 +/- 0.05 84.80 +/- 0.55 F05 82.77 +/- 0.00 243.26 55.98 +/- 0.11 66.86 +/- 0.08 81.13 +/- 0.15 F01 F02 F03 F04 F05 Thermal unfolding Unfolding-induced aggregation Figure 5: Analysis of mAb stability by nanoDSF with backreflection aggregation detection. Aggregation detection of the samples shown in Figure 4. Aggregation was monitored in parallel to the intrinsic fluorescence. T agg values represent averages with standard deviation from triplicate measurements, the degree of aggregation at 95 °C represent averages from triplicate measurements.