Download Studying the binding affinity

Studying the binding affinity
of different ERK2 binding partners
with MicroScale Thermophoresis
Clemens Entzian, Lukas Kniep, Dr. Thomas Schubert,
2bind GmbH
Antibody 1B3B9 (Merck)
Molecular Size
ERK2 is an important player in multiple
biochemical signaling pathways and is
involved in different cancer types. This highly
relevant pharmaceutical target represents an
optimal model system to demonstrate the
capabilities of our MicroScale Thermophoresis
(MST) platform.
fraction bound
1
0.8
0.6
150 kDa
0.4
0.2
0
0.1 1 10 100 1000 104
concentration titrant/nM
Kd = 137.9 + 23.3 nM
Aptamer C5.71
The results (figure to the right) demonstrate
that our MST platform is perfectly suited to
study molecular interactions independent of
the size and mass of the ligand.
Therefore, MicroScale Thermophoresis is the
optimal tool to characterize drug-target,
antibody-antigen
or
aptamer-target
interactions in terms of basic binding
parameters.
0.4
3 kDa
0.2
0
0.1 1
10 100 1000 104
Erk Activation
Inhibitor Peptide I (Merck)
1.2
fraction bound
• ERK2 Inhibitor I (Merck)
• ERK2 Activation Inhibitor
Peptide I (Merck)
• Aptamer C5.71 (Mayer lab, Bonn)
• Antibody 1B3B9 (Merck)
0.6
concentration titrant/nM
Kd = 171.1 + 36.0 nM
0.8
1 kDa
0.4
0
10 100 1000 104 105 106
concentration titrant/nM
Kd = 4854.2 + 1686 nM
ERK2 Inhibitor I (Merck)
1.2
fraction bound
In this example study, the binding affinity of
ERK2 (MAPK1, mitogen-activated protein
kinase 1) towards the following four different
binding partners (size range from 0.3 kDa to
150 kDa) was studied by MST:
fraction bound
1
0.8
0.8
0.4
0.3 kDa
0
1 10 100 1000 104 105 106
concentration titrant/nM
Kd = 2025 + 924 nM
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