Table 1.
Chelate:protein substitution levels under different reaction conditions for NOTA conjugation of pertuzumab F(ab')2.
| F(ab')2 protein concentration (mg/mL) |
||||||
|---|---|---|---|---|---|---|
| 2.5 | 5.0 | 10.0 | ||||
| Molar ratio of NOTA:F(ab')2 |
ITLC-SG* |
UV assay† |
ITLC-SG |
UV assay |
ITLC-SG |
UV assay |
| 5:1 | 2.1 ± 0.0 | 2.8 ± 1.1 | 3.8 ± 0.4 | 3.7 ± 1.7 | 4.3 ± 2.5 | 5.8 ± 2.9 |
| 10:1 | 4.1 ± 1.9 | 4.2 ± 1.4 | 6.9 ± 1.8 | 5.0 ± 1.1 | 6.5 ± 1.8 | 9.9 ± 8.8 |
Values shown are mean ± SD (n = 3–8).
*
Instant thin layer-silica gel chromatography developed in 0.1 M sodium citrate, pH 5.0.
†
Measurement of absorbance at 280 nm with subtraction of the contribution from pertuzumab F(ab')2.