Peroxide Residual and Oxidation of mAb1 Formulation (10 mg/mL) Spiked with Various Concentrations of H2O2 at 5 °C Storage
| Assay | Time (months) | H2O2 Concentration (ng/mL) | |||||
|---|---|---|---|---|---|---|---|
| 0 (control) | 10 | 50 | 100 | 250 | 1000 | ||
| Amplex® UltraRed HRP Assay—Peroxide (ng/mL) | 0 | <LOD | 12 | 47 | 100 | 254 | 998 |
| 3 | <LOD | <LOD | 18 | 43 | 111 | 466 | |
| 6 | <LOD | <LOD | <LOD | 19 | 59 | 268 | |
| 9 | <LOD | <LOD | <LOD | 6 | 23 | 133 | |
| 12 | <LOD | <LOD | <LOD | <LOD | 8 | 63 | |
| 18 | <LOD | <LOD | <LOD | <LOD | <LOD | 13 | |
| 24 | <LOD | <LOD | <LOD | <LOD | <LOD | <LOD | |
| Peptide Mapping—HPLC (Oxidation Detected) | 0 | N | NT | NT | N | NT | NT |
| 3 | N | N | N | N | N | Y | |
| 6 | N | N | N | N | Y | Y | |
| 9 | N | N | N | N | N | Y | |
| 12 | N | N | N | N | N | NT | |
| 18 | N | N | N | N | N | NT | |
| 24 | N | N | N | N | N | NT | |
HPLC, high-performance liquid chromatography; HRP, horseradish peroxidase; LOD, limit of detection; N, no oxidation detected: peptide map profile of spike sample was comparable to the control; NT, not tested; Y, oxidation detected: new peaks were observed in peptide map profile of spike sample when compared with control.