Table 1 Properties of eGFP-binding DARPins

From: Molecular features defining the efficiency of bioPROTACs

Name

Original Name

KD (Anisotropy) [nM]

KD (SPR) [nM]

kon (SPR)[M−1 s−1 106]

koff(SPR) [s−1 10−4]

DM (GdmCl) [M] n = 2

Tm [°C]

DP1

3G190.24

nd

2.76

2.56

70.8

3.3

73

DP2

2G156

nd

0.235

6.62

15.6

3.2

71

DP3

3G86.32

<5 (Ref. 22)

1.38

0.68

9.4

2.8

69

DP4

2G71

nd

0.108

48.4

52.4

3.1

65

DP5

3G168

<5 (Ref. 22)

0.682

0.929

6.34

3.7

>90 °C

DP6

3G124

<5 (Ref. 22)

0.271

2.09

5.67

4.3

>90 °C

DP6dest

–

nd

3.94

1.53

60.3

0.9

nd

DP6W112A

–

25,210 ± 10,000

477

nd

nd

4.8

>90 °C

DP6lysfree

–

nd

1.07

3.76

0.353

nd

87

DP7

3G146

669 ± 156

nd

nd

nd

3.6

81

DP8

3G86.1

<5 (Ref. 22)

1.34

0.767

10.2

3.8

>90 °C

DP9

3G61

76 ± 21 (Ref. 22)

0.376

3.42

12.9

2.4

57

  1. Kinetics were determined by SPR spectroscopy (Supplementary Fig. S19). KD values of select DARPins were determined by fluorescence anisotropy (Supplementary Fig. S20). Denaturation midpoints (DM) were determined by guanidinium chloride unfolding measured with CD-spectroscopy (Supplementary Fig. S21). Melting temperatures as indicated were determined by SYPRO orange dye binding on a qPCR cycler (Supplementary Fig. S23).
  2. nd not determined.
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