Activity of representative nanobodies: off-rate determinations on biotinylated CXCR2 1–19 peptide (single experiment using periplasmic extracts) and inhibition of labeled CXCL1 binding to CHO-CXCR2 cells (purified nanobodies)
Number is given in parentheses.
| Nanobody | 1–19 Peptide Binding | CXCL1 Binding Assay | |
|---|---|---|---|
| kd [1/s] | pIC50 | % Maximal Inhibition | |
| 2B2 | 7.4 × 10−3 | 8.87 ± 0.14 (6)a | 40.3 ± 2.4a |
| 127D1 | 7.7 × 10−4 | 9.40 ± 0.11 (4)b | 59.8 ± 2.0b |
| 54B12 | 1.4 × 10−3 | 9.14 ± 0.25 (2)a,b | 35.0 ± 5.0a |
| 97A9 | No binding | 7.85 ± 0.06 (8)c | 93.2 ± 2.4c |
| 163E3 | No binding | 8.02 ± 0.02 (4)c,d | 94.0 ± 1.4c |
| 163D2 | No binding | 8.31 ± 0.10 (5)d | 96.8 ±1.1c |
Values annotated with the same superscript letter(s) are not statistically significantly different, based on comparison of all nanobodies for the CXCL1-binding assay data (P < 0.05; Tukey’s honest significant difference test after one-way analysis of variance).