©2021 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved.
6
APPLICATION NOTE
Figure 2: Profiles of two "groups" within each group structure. A is Protein A (non-mutated) + Protein A1-4 (mutants). B is Protein G (non-mutated) + Protein G1-4
(mutants). Each mutant represents only 1-2 site mutations in a non-antigen-binding region of the IgG-derived mAb. Note the overall profile shapes remain similar for DSF
(top), turbidity (centre), and DLS (bottom), though the mutations have a noticeable effect on the structural profile.
Ratio
350
nm
/
330
nm
Turbidity
[mAU]
Cumulant
Radius
[nm]
Temperature [ºC]
A B
Ratio
350
nm
/
330
nm
Turbidity
[mAU]
Cumulant
Radius
[nm]
Temperature [ºC]
Protein A
Protein A1
Protein A2
Protein A3
Protein A4
Protein G Protein G2
Protein G1 Protein G3
Protein G4
T
m1
T
on
T
turb
T
scattering
PDI (25ᵒC)
Protein A 64.99 60.48 64.33 63.42 0.1
Protein A1 60.69 56.93 60.47 60.72 0.08
Protein A2 63.69 59.57 64.86 63.38 0.77
Protein A3 60.08 54.91 71.91 58.96 0.02
Protein A4 60.63 56.40 61.29 60.71 1.23
Protein G 71.47 65.26 82.97 82.34 0.03
Protein G1 69.31 54.70 83.81 69.07 0.04
Protein G2 68.76 59.99 83.81 69.05 0.30
Protein G3 69.69 63.52 81.20 69.04 0.02
Protein G4 72.27 63.52 83.81 72.55 0.64
Table 2: Summary data of first unfolding
temperature, unfolding onset, turbidity
onset, scattering onset, and PDI for each
group member for two representative groups.
Non-mutated in bold. A PDI <0.1 is considered
highly monodisperse; anything above 0.25
is considered highly polydisperse -- high PDI
values highlighted in red.
