Table 2.
Characterization of ProC6C.
| Test | Method | 1L consistency 1 | 1L consistency 2 | 1L consistency 3 | 5 L lab-scale |
|---|---|---|---|---|---|
| pH | pH meter | 8.0 | 8.0 | 8.0 | 8.0 |
| Color and appearance | Visual observation | Colorless | Colorless | Colorless | Colorless |
| Endotoxin | LAL | 11 EU/mg | 9.0 EU/mg | 12.5 EU/mg | 17 EU/mg |
| Residual host-cell protein by ELISA | Immunoenzymatic Assay | <1% | <1% | <1% | <1% |
| Protein content | A280/Nanodrop | 0.65 mg/mL | 0.58 mg/mL | 0.60 mg/mL | 0.8 mg/mL |
| Accurate quantification of the total protein | AAA analysis | 0.68 mg/ml | ND | ND | 0.81 mg/ml |
| Identity and integrity (SDS-PAGE) | SDS-PAGE both nonreduced and reduced | Major monomer band | Major monomer band | Major monomer band | Major monomer band |
| Identity (WB) | Both nonreduced and reduced Western Blot against monoclonal antibodies 45.1 | Major monomer band | Major monomer band | Major monomer band | Major monomer band |
| Purity RP-HPLC | RP-HPLC | 98.3% | >97.8% | >96.9% | >97.3% |
| Identity, integrity and purity (SE-HPLC) | SE-HPLC | 97.3% monomer | 98.5% monomer | 98.0% monomer | 96.5% monomer |
| Intact mass spectrometry analysis of full-length protein | Outsourced: Intact mass spec (LC–MS/MS) under nonreducing and reducing conditions | Nonreduced (36308.54 Da) Reduced (36314.599 Da) | ND | ND | Nonreduced (36308.54 Da) Reduced (36314.599 Da) |