Application Notes

7 APPLICATION NOTE ©2017 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved. Figure 5: Formulation screening of PPA. In order to find optimal conditions for an increased thermal stability of PPA, its thermal unfolding was monitored under 16 different additive conditions. A clear shi of T m towards higher values is visible in the fluorescence ratio plots for each additive. Quantification of T m under different conditions renders the addition of 30 % trehalose most effective, while glycerol has the weakest effect. 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 0 10 20 30 40 54 56 58 60 62 64 66 68 Trehalose Glycerol Sucrose Sorbitol Sorbitol (% w/v) 0 10 20 30 40 Sucrose (% w/v) 0 10 20 30 40 Glycerol (% w/v) 0 10 20 30 40 Trehalose (% w/v) 0 10 20 30 40 Pig pancreatic a-amylase Pig pancreatic a-amylase A B % additive (w/v) T (°C) m Temp (°C) Temp (°C) Temp (°C) Temp (°C) 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 0 10 20 30 40 54 56 58 60 62 64 66 68 Trehalose Glycerol Sucrose Sorbitol Sorbitol (% w/v) 0 10 20 30 40 Sucrose (% w/v) 0 10 20 30 40 Glycerol (% w/v) 0 10 20 30 40 Trehalose (% w/v) 0 10 20 30 40 Pig pancreatic a-amylase Pig pancreatic a-amylase A B % additive (w/v) T (°C) m Temp (°C) Temp (°C) Temp (°C) Temp (°C) 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 Temp (°C) Temp (°C) 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 0 10 20 30 40 54 56 58 60 62 64 66 68 Trehalose Glycerol Sucrose Sorbitol Sorbitol (% w/v) 0 10 20 30 40 Sucrose (% w/v) 0 10 20 30 40 Glycerol (% w/v) 0 10 20 30 40 Trehalose (% w/v) 0 10 20 30 40 Pig pancreatic a-amylase Pig pancreatic a-amylase A B % additive (w/v) T (°C) m Temp (°C) Temp (°C) Temp (°C) Temp (°C) 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 0 10 20 30 40 54 56 58 60 62 64 66 68 Trehalose Glycerol Sucrose Sorbitol Sorbitol (% w/v) 0 10 20 30 40 Sucrose (% w/v) 0 10 20 30 40 Glycerol (% w/v) 0 10 20 30 40 Trehalose (% w/v) 0 10 20 30 40 Pig pancreatic a-amylase Pig pancreatic a-amylase A B % additive (w/v) T (°C) m Temp (°C) Temp (°C) Temp (°C) Temp (°C) 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 Temp (°C) Temp (°C) Temp (°C) A Pig pancreatic α-amylase 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 20 30 40 50 60 70 80 90 0 10 20 30 40 54 56 58 60 62 64 66 68 Trehalose Glycerol Sucrose Sorbitol Sorbitol (% w/v) 0 10 20 30 40 Sucrose (% w/v) 0 10 20 30 40 Glycerol (% w/v) 0 10 20 30 40 Trehalose (% w/v) 0 10 20 30 40 Pig pancreatic a-amylase Pig pancreatic a-amylase A B % additive (w/v) T (°C) m Temp (°C) Temp (°C) Temp (°C) Temp (°C) 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 1.20 1.25 1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 F350 / F330 % additive (w/v) B Pig pancreatic α-amylase

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