Table 6.
Binding affinities for triple-thiol rHSA variants toward hFcRn, pH 5.5
KD (koff/kon) is shown as mean ± S.D. Average Biolayer Interferometry data from three replicates are given, from two-fold dilution series from 3 to 0.1875 μm albumin. (For binding curves, see Fig. 2.) NA, not available.
| Albumin | KD average | Kon average | Koff average |
|---|---|---|---|
| μm | m−1 s−1 | s−1 | |
| Single-thiol WT | 0.55 ± 0.20 | 1.38 × 104 ± 1.69 × 103 | 7.76 × 10−3 ± 2.50 × 10−3 |
| Triple-thiol WT | 0.25 ± 0.05 | 1.79 × 104 ± 2.70 × 103 | 4.46 × 10−3 ± 1.48 × 10−3 |
| Triple-thiol WT-3xAF680 | 0.88 ± 0.01 | 7.57 × 103 ± 1.82 × 103 | 6.45 × 10−3 ± 5.52 × 10−4 |
| Triple-thiol NB | NA | NA | NA |
| Triple-thiol NB -3xAF680 | NA | NA | NA |
| Triple-thiol HBII | 0.02 ± 0.006 | 2.01 × 104 ± 4.13 × 103 | 3.08 × 10−4 ± 9.96 × 10−5 |
| Triple-thiol HBII -3xAF680 | 0.04 ± 0.017 | 1.25 × 104 ± 1.70 × 103 | 4.53 × 10−4 ± 2.33 × 10−4 |