A designed ankyrin repeat protein evolved to picomolar affinity to Her2

A designed ankyrin repeat protein evolved to picomolar affinity to Her2

A designed ankyrin repeat protein evolved to picomolar affinity to Her2
Christian Zahnd, Emanuel Wyler, Jochen M. Schwenk, Daniel Steiner, Michael C. Lawrence,
Neil M. McKern, Frédéric Pecorari, Colin W. Ward, Thomas O. Joos and Andreas Plückthun
Supplementary Figure 1. The affinities of different DARPins were determined by surface
Plasmon resonance (SPR) using a Biacore 3000 instrument. Her2 ECD was immobilized on a
flow cell at low concentrations. Binding of the DARPin to the immobilized target protein was
compared to binding to an empty flow cell at different concentrations of the DARPin. Data
were processed by the software Scrubber and evaluated by global fitting using the software
CLAMP32. Each DARPin was injected several times at different concentrations. Note that H3HAVD was injected at significantly higher concentrations than the other DARPins. DARPins
G3-D and G3-A were injected at 20 nM, 50 nM, 100 nM and 150 nM, G3-AVD was injected
at 50 nM, 100 nM, 200 nM and 300 nM, G3-HAVD was injected at 500 nM, 1000 nM,
2000 nM, 4000 nM, 6000 nM and 8000 nM, H10-2-G5 was injected at 1 nM, 5 nM, 10 nM,
20 nM, 30 nM, 50 nM, 75 nM, 100 nM and 150 nM, H10-2-D11 was injected at 10 nM,
20 nM, 30 nM, 50 nM, 75 nM, 100 nM and 150 nM, H10-2-D12 was injected at 0 nM,
100 nM, 200 nM, 300 nM and 400 nM and H10-2-A2 was injected at 0 nM, 1 nM, 5 nM,
10 nM, 20 nM, 30 nM, 50 nM, 100 nM and 150 nM.
Supplementary Figure 1
30
25
G3-HAVD
Resonance Unit (RU)
Resonance Unit (RU)
25
20
15
10
5
15
10
5
0
0
G3-AVD
20
15
10
5
0
H10-2-D11
30
Resonance Unit (RU)
Resonance Unit (RU)
H10-2-D12
20
20
10
0
G3-A
H10-2-A2
Resonance Unit (RU)
Resonance Unit (RU)
30
20
10
15
10
5
0
0
G3-D
20
10
0
H10-2-G5
30
Resonance Unit (RU)
30
Resonance Unit (RU)
20
20
10
0
0
200
400
time (s)
600
800
0
200
400
time (s)
600
800