DS–IMP and PQ Specification Examples for Typical Analytical Methods
| Test/Specification | IMP Phase 1–2 | IMP Pivotal or Phase 3 | PQ Lots (PV Stage 2) | Reported Results Example |
|---|---|---|---|---|
| Appearance | Clarity: Clear to opalescent Color: Colorless to yellow liquid, Particles: Free from or practically free from visible particles | Clarity: NMT EP RS III Color: NMT Y5 Particles: Free from or practically free from visible particles | Clarity: NMT EP RS III Color: NMT Y5 Particles: Free from or practically free from visible particles | Clarity: RS I Color: Y7 Particles: Free from visible particles |
| Total protein | Nominal value ± 10.0% | Nominal value ± 10.0% | Nominal value ± 10.0% | 1.05 mg/mL 10.2 mg/mL 52.0 mg/mL 103 mg/mL |
| Capillary isoelectric focusing(cIEF) | Peak pattern consistent with Reference Standard; report % monomer, % total acidic peaks, % total basic peaks | Peak pattern consistent with Reference Standard; monomer NLT 50%, report total % acidic peaks, report % total basic peaks | Peak pattern consistent with Reference Standard; monomer: NLT 65%, total acidic peaks: NMT 30% | Peak pattern consistent with Reference Standard; monomer: 72%, total acidic peaks: 14% |
| Target Binding Bioassay | 50–150% (symmetrical) of Reference Standard binding 60–167% (geometrical) of Reference Standard binding | N/A | N/A | 98% |
| Mode-of-Action (MOA)-simulated Bioassay | N/A | 60–140% (symmetrical) of Reference Standard binding 70–143% (geometrical) of Reference Standard binding | 90–120% (symmetrical) of Reference Standard binding 90–125% (geometrical) of Reference Standard binding | 98% |
| Reducing gel electrophoresis | Electrophoretic peak pattern consistent with Reference Standard; area percent purity of heavy + light chains: NLT 95.0%; total area percent of impurities: NMT 5.0% | Area percent purity of heavy + light chains: NLT 97.0%; total area percent of impurities: NMT 3.0% | Area percent purity of heavy + light chains: NLT 98.5%; total area percent of impurities: NMT 1.5% | Area percent purity of heavy + light chains: 99.2%; total area percent of impurities: 0.8% |
| Non-reducing gel electrophoresis | Electrophoretic peak pattern consistent with Reference Standard; major product peak: NLT 95%; total area percent of impurities: NMT 5.0% | Major product peak: NLT 97.0%; total area percent of impurities: NMT 3.0% | NLT 98.5%; total area percent of impurities: NMT 1.5% | Major product peak: 99.2%; total area percent of impurities: 0.8% |
| HPSEC | Major product peak: NLT 95.0%; report % aggregates; report % fragments | Major product peak: NLT 96.0%; aggregates: NMT 4.0%; fragments: NMT 4.0% | Major product peak: NLT 98.3%; aggregates: NMT 1.7%; fragments: NMT 1.7% | Major product peak: 99.2.0%; aggregates: 0.5%; fragments: 0.3% |
| Host cell DNA | LT 0.10 ng DNA/mg protein | LT 0.10 ng DNA/mg protein | LT 20 pg DNA/mg protein | 0.02 ng DNA/mg protein 2 pg DNA/mg protein |
| CHO Host cell protein | NMT 100 ng/mg protein | NMT 100 ng/mg protein | NMT 20 ng/mg protein | 3 ng/mg protein |
| Protein A | NMT 50 ng/mg protein | NMT 50 ng/mg protein | NMT 10 ng/mg protein | 2 ng/mg protein |
| Bioburden | NMT 1 CFU per 10 mL | NMT 1 CFU per 10 mL | NMT 1 CFU per 10 mL | 0 CFU per 10 mL |
| Endotoxin (LAL) | NMT 0.30 EU/mg protein | NMT 0.30 EU/mg protein | NMT 0.10 EU/mg protein | 0.01 EU/mg protein |