Table 3.
Characterization of R0.6C.
| Test | Method | Result | |||
|---|---|---|---|---|---|
| Consistency 1(1L) | Consistency 2(1L) | Consistency 3(1L) | Lab-Scale(5L) | ||
| pH | pH meter | 8.0 | 8.0 | 8.0 | 8.0 |
| Color and Appearance | Visual observation | Colorless | Colorless | Colorless | Colorless |
| Molar Extinction Coefficient (M-1•cm-1) | spectroscopy | 13, 913.4a | NDb | ||
| Protein Content | A280/Nanodrop | 0.7 mg/ml | 0.8 mg/ml | 0.75 mg/ml | 1.1 mg/ml |
| Accurate quantification of the total protein | AAA analysis | 0.72 mg/ml | ND | ND | 1.2 mg/ml |
| Endotoxin | LAL | 17 EU/mg | 8.0 EU/mg | 11 EU/mg | 6.0 EU/mg |
| Residual host-cell DNA | Qubit Fluorescence Assay | 2.7 pg/µla | |||
| N-terminal sequencing analysis | N-terminal sequencing | AERSTSENRNa | NDb | ||
| Residual host-cell protein by ELISA | Immuno-enzymatic Assay | <1% | <1% | <1% | <1% |
| Identity and Integrity (SDS-PAGE) |
SDS-PAGE both non-reduced and reduced | Major monomer band | Major monomer band | Major monomer band | Major monomer band |
| Identity (WB) |
Non-reduced 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 | 97.6% | >97.3% | >96.9% | >95% |
| Identity, Integrity and Purity (SE-HPLC) |
SE-HPLC | 98.5% monomer | 97.9% monomer | 98.2% monomer | 95.5% monomer |
| Intact Mass spectrometry analysis of full-length protein (LC-MS/MS) |
Intact mass spec under non-reducing and reducing conditions | Non-reduced (70257.906Da) a
Reduced (70263.852Da) a |
Non-reduced (70257.916Da) Reduced (70263.863Da) |
||
aA single sample from the three runs were utilized for contracted testing. Consistency run#1 solely analyzed.
bND, Not determined or performed on sample.