TECHNICAL NOTE
Tycho is a trademark
of NanoTemper Technologies GmbH, Munich, Germany. NanoTemper is a registered trademark of NanoTemper Technologies GmbH, Munich,
Germany. ©2017 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved. TE-TY-002-01
nanotempertech.com
Figure 2: Label-free thermal shi analysis of. 10 µM maltose-binding protein
(MBP) interacting with 100 µM maltose using Tycho NT.6. Dramatic differences in
the initial ratio values are likely due to the conformational changes of MBP upon
binding of maltose. Vertical lines indicate inflection temperatures (T
i
), colored
circles at the y-axis indicate initial ratio values.
Figure 3: Testing the interaction of bacterial cell wall complex (NAG) with
lyso
zyme using the Tycho
NT.6. 5 µM lysozyme was incubated with 50 µM 3'NAG
and analyzed. Vertical lines indicate inflection temperatures (T
i
), colored circles
at the y-axis indicate initial ratio values.
30 40 50 60 70 80 90 100
0.90
0.95
1.00
1.05
1.10
1.15
1.20
Ratio
350
nm
/
330
nm
Temperature (°C)
MBP
MBP + 100 µM maltose
0.6
0.7
0.8
0.9
1.0
1.1
1.2
1.3
30 40 50 60 70 80 90 100
Temperature (°C)
Lysozyme
Lysozyme + 50 µM 3'NAG
Ratio
350
nm
/
330
nm
Conclusions
These examples illustrate that the Tycho NT.6 system
can provide a fast and reliable yes/no binding checks
utilizing label-free thermal shi assays. In addition,
interactions detected on the system can serve as
an initial test or predictor of functionality. Proteins
interacting with their ligands as expected will likely
have appropriate functionality in further downstream
analysis to be run. Experiments on the Tycho NT.6 are
fast to perform, easy to carry out, utilize microliter
amounts of sample and serve as a standard quality
validation tool for scientists doing protein research.
Reference
1. Telmer, P.G. and B.H. Shilton, Insights into the conformational
equilibria of maltose-binding protein by analysis of high affinity
mutants. J Biol Chem, 278(36): p. 34555-67 (2003).
