Table 2.
Summary of results for physicochemical similarity assessment between CT-P10, EU-rituximab and US-rituximab.
| Min – Max Range (Mean ± SD) |
||||||
|---|---|---|---|---|---|---|
| Quality Attribute / Test Method | EU-Rituximab (N = 15) | CT-P10 Drug Product (N = 15) | US-Rituximab (N = 15) | |||
| Primary/Higher Order Structure | Molar Absorptivity | Molar Absorptivity (L∙mol−1∙cm−1) | 186,065 – 229,830 (212,656 ± 11,707) | 193,519 – 235,223 (213,636 ± 11,239) | 196,166 – 224,104 (212,717 ± 7,487) | |
| Extinction Coefficient (L∙g−1∙cm−1) | 1.29 – 1.59 (1.47 ± 0.08) | 1.34 – 1.63 (1.48 ± 0.08) | 1.36 – 1.55 (1.47 ± 0.05) | |||
| Peptide Mapping (LC-MS) | Deamidation (%) | HC Asn55 | 1.6 – 2.3 (1.9 ± 0.2) | 1.7 – 2.1 (1.9 ± 0.1) | 1.6 – 2.1 (1.9 ± 0.1) | |
| HC Asn290 | 0.1 – 0.2 (0.2 ± 0.0) | 0.1 – 0.2 (0.2 ± 0.0) | 0.1 – 0.2 (0.2 ± 0.0) | |||
| HC Asn319 | 0.4 – 0.6 (0.5 ± 0.1) | 0.5 – 0.7 (0.5 ± 0.1) | 0.4 – 0.6 (0.5 ± 0.1) | |||
| HC Asn365 | 0.1 – 0.2 (0.1 ± 0.0) | 0.1 – 0.2 (0.1 ± 0.0) | 0.1 – 0.2 (0.1 ± 0.0) | |||
| HC Asn388 | 1.8 – 2.5 (2.1 ± 0.2) | 1.5 – 2.1 (1.8 ± 0.2) | 1.8 – 2.3 (2.0 ± 0.2) | |||
| LC Asn136 | 0.1 – 0.1 (0.1 ± 0.0) | 0.1 – 0.1 (0.1 ± 0.0) | 0.1 – 0.1 (0.1 ± 0.0) | |||
| Oxidation (%) | HC Met34 | 0.1 – 0.2 (0.2 ± 0.1) | 0.1 – 0.2 (0.1 ± 0.1) | 0.1 – 0.2 (0.2 ± 0.1) | ||
| HC Met81 | 0.3 – 0.6 (0.4 ± 0.1) | 0.3 – 0.5 (0.4 ± 0.1) | 0.3 – 0.6 (0.4 ± 0.1) | |||
| HC Met256 | 1.9 – 2.9 (2.4 ± 0.3) | 2.0 – 3.6 (2.5 ± 0.5) | 1.9 – 3.4 (2.5 ± 0.4) | |||
| HC Met432 | 0.5 – 0.8 (0.6 ± 0.1) | 0.4 – 1.1 (0.7 ± 0.2) | 0.5 – 1.0 (0.7 ± 0.2) | |||
| LC Met21 | 0.1 – 0.3 (0.2 ± 0.1) | 0.1 – 0.4 (0.2 ± 0.1) | 0.1 – 0.3 (0.2 ± 0.1) | |||
| N-terminal pyro-Glu HC (%) | 99.7 – 99.9 (99.8 ± 0.1) | 99.5 – 99.8 (99.7 ± 0.1) | 99.6 – 99.9 (99.8 ± 0.1) | |||
| N-terminal pyro-Glu LC (%) | 96.1 – 98.9 (97.6 ± 0.7) | 97.1 – 98.7 (95.7 ± 0.5) | 97.4 – 98.7 (97.4 ± 0.9) | |||
| C-terminal Lys truncation HC (%) | 97.6 – 98.4 (98.1 ± 0.3) | 97.1 – 98.7 (98.1 ± 0.4) | 97.4 – 98.7 (98.2 ± 0.3) | |||
| Intact Mass (LC-MS) | G0F-G0F (Da) | 147085.9 – 147087.9 (147087.0 ± 0.6) | 147085.3 – 147086.5 (147085.8 ± 0.4) | 147085.9 – 147.088.3 (147087.0 ± 0.7) | ||
| G0F-G1F (Da) | 147246.9 – 147.249.4 (147248.3 ± 0.8) | 147246.9 – 147248.2 (147247.5 ± 0.4) | 147246.9 – 147249.3 (147248.4 ± 0.7) | |||
| G1F-G1F or G0F-G2F (Da) | 147408.2 – 147411.3 (147409.8 ± 0.8) | 147408.3 – 147410.3 (147408.9 ± 0.5) | 147408.5 – 147411.0 (147409.8 ± 0.7) | |||
| G1F-G2F (Da) | 147569.6 – 147574.1 (147571.4 ± 1.1) | 147568.8 – 147571.2 (147570.0 ± 0.7) | 147570.1 – 147573.9 (147571.6 ± 1.2) | |||
| Free Thiol Analysis | Average (free SH/IgG, µM/µM) | 0.30 – 0.35 (0.32 ± 0.02) | 0.27 – 0.31 (0.29 ± 0.01) | 0.28 – 0.36 (0.32 ± 0.02) | ||
| FTIR (cm−1) | Amide I | 1641 – 1642 (1641 ± 0.1) | 1641 – 1641 (1641 ± 0.1) | 1641 – 1641 (1641 ± 0.1) | ||
| Amide II | 1527 – 1529 (1528 ± 0.4) | 1527 – 1529 (1528 ± 0.4) | 1528 – 1528 (1528 ± 0.2) | |||
| DSC (°C) | Tm1 | 71.8 – 71.9 (71.9 ± 0.0) | 71.9 – 72.0 (71.9 ± 0.0) | 71.8 – 72.0 (71.9 ± 0.0) | ||
| Tm2 | 76.1 – 76.1 (76.1 ± 0.0) | 76.1 – 76.1 (76.1 ± 0.0) | 76.1 – 76.1 (76.1 ± 0.0 | |||
| Tm3 | 83.2 – 83.4 (83.3 ± 0.1) | 83.2 – 83.3 (83.3 ± 0.0) | 83.2 – 83.4 (83.3 ± 0.1) | |||
| Content | Protein Concentration (UV280) (mg/mL) | 10.0 – 10.6 (10.4 ± 0.2) | 9.7 – 10.6 (10.1 ± 0.3) | 10.0 – 10.5 (10.2 ± 0.2) | ||
| Purity/Impurity | SEC-HPLC | Monomer (%) | 98.75 – 99.21 (99.01 ± 0.15) | 99.17 – 99.50 (99.30 ± 0.10) | 98.60 – 99.06 (98.81 ± 0.17) | |
| HMW (%) | 0.73 – 1.13 (0.89 ± 0.13) | 0.26 – 0.82 (0.53 ± 0.19) | 0.86 – 1.27 (1.07 ± 0.14) | |||
| LMW (%) | 0.07 – 0.14 (0.10 ± 0.02) | 0.00 – 0.47 (0.17 ± 0.17) | 0.08 – 0.16 (0.12 ± 0.03) | |||
| SEC-MALS | Monomer (MW, kDa) | 137 – 139 (138 ± 1) | 137 – 139 (138 ± 1) | 137 – 139 (138 ± 1) | ||
| HMW (MW, kDa) | 247 – 344 (293 ± 25) | 166 – 307 (197 ± 40) | 237 – 364 (303 ± 35) | |||
| AUC | Monomer (%) | 97.88 – 98.48 (98.19 ± 0.23) | 97.50 – 98.52 (98.18 ± 0.30) | 97.04 – 98.45 (98.05 ± 0.40) | ||
| Dimer (%) | 1.53 – 2.12 (1.81 ± 0.23) | 1.48 – 2.50 (1.82 ± 0.30) | 1.55 – 2.96 (1.95 ± 0.40) | |||
| Monomer (s-value) | 6.03 – 6.28 (6.18 ± 0.06) | 6.10 – 6.32 (6.18 ± 0.05) | 6.09 – 6.25 (6.17 ± 0.07) | |||
| Dimer (s-value) | 8.78 – 9.12 (8.97 ± 0.11) | 8.70 – 9.19 (8.96 ± 0.12) | 8.60 – 9.09 (8.95 ± 0.12) | |||
| CE-SDS | NR | Intact IgG (%) | 90.61 – 96.16 (93.21 ± 1.44) | 94.54 – 97.18 (95.78 ± 0. 77) | 91.33 – 95.89 (93.40 ± 1.18) | |
| R | H+L (%) | 99.09 – 99.71 (99.53 ± 0.16) | 98.97 – 99.34 (99.20 ± 0.11) | 99.46 – 99.70 (99.57 ± 0.08) | ||
| Charge variants | IEC-HPLC | Acidic Group (Peak 1+2+3) (%) | 18.7 – 22.8 (20.8 ± 1.2) | 15.6 – 18.5 (16.7 ± 0.7) | 19.0 – 22.5 (20.9 ± 1.0) | |
| Main Peak (Peak 4) (%) | 66.5 – 72.8 (70.4 ± 1.8) | 65.5 – 68.4 (67.1 ± 0.9) | 67.6 – 72.0 (69.9 ± 1.5) | |||
| Basic Group (Peak 5+6+7) (%) | 7.0 – 11.0 (8.8 ± 1.0) | 15.4 – 17.4 (16.3 ± 0.5) | 8.0 – 10.6 (9.2 ± 0.9) | |||
| Glycosylation | Oligosaccharide Profile | Galactosylation (%) | 44.59 – 56.48 (51.07 ± 3.57) | 47.89 – 49.70 (48.63 ± 0.48) | 47.01 – 54.63 (50.78 ± 2.28) | |
| Afucosylation (%) | 0.77 – 2.05 (1.21 ± 0.33) | 0.36 – 0.93 (0.55 ± 0.14) | 0.66 – 1.89 (1.16 ± 0.36) | |||
| High Mannose (%) | 1.22 – 1.88 (1.43 ± 0.16) | 2.97 – 3.50 (3.29 ± 0.15) | 1.25 – 2.19 (1.63 ± 0.24) | |||
| Total Afucosylation (%) | 1.99 – 3.47 (2.64 ± 0.40) | 3.33 – 4.41 (3.84 ± 0.25) | 2.02 – 3.44 (2.79 ± 0.35) | |||
| Sialylation (%) | 0.87 – 2.98 (1.90 ± 0.63) | 1.34 – 1.50 (1.41 ± 0.05) | 0.84 – 2.93 (1.91 ± 0.64) | |||
| N-linked Glycan Analysis | Galactosylation (%) | 44.70 – 56.51 (51.17 ± 3.07) | 46.74 – 51.26 (48.89 ± 1.38) | 46.17 – 56.08 (50.97 ± 3.2) | ||
| Afucosylation (%) | 0.46 – 1.34 (0.80 ± 0.22) | 0.23 – 0.54 (0.36 ± 0.09) | 0.40 – 1.24 (0.78 ± 0.24) | |||
| High Mannose (%) | 1.46 – 2.48 (1.79 ± 0.27) | 3.13 – 4.09 (3.62 ± 0.27) | 1.54 – 2.38 (1.83 ± 0.26) | |||
| Total Afucosylation (%) | 2.10 – 3.09 (2.59 ± 0.30) | 3.46 – 4.62 (3.98 ± 0.32) | 2.22 – 3.03 (2.62 ± 0.24) | |||
| Sialylation (%) | 0.95 – 1.76 (1.44 ± 0.23) | 0.80 – 1.16 (0.92 ± 0.09) | 0.93 – 2.16 (1.54 ± 0.33) | |||
| Sialic Acid Analysis | Molar Ratio (sialic acid / protein mol / mol) | 0.07 – 0.12 (0.10 ± 0.01) | 0.07 – 0.09 (0.08 ± 0.01) | 0.07 – 0.14 (0.10 ± 0.02) | ||
| Monosaccharide Analysis (Molar Ratio) | Fuc | 1.1 – 1.3 (1.2 ± 0.1) | 1.1 – 1.3 (1.2 ± 0.1) | 1.1 – 1.4 (1.2 ± 0.1) | ||
| GlcNAc | 5.1 – 5.9 (5.6 ± 0.2) | 5.2 – 5.9 (5.5 ± 0.2) | 5.1 – 6.4 (5.6 ± 0.3) | |||
| Gal | 0.8 – 1.2 (0.9 ± 0.1) | 0.7 – 0.8 (0.8 ± 0.0) | 0.7 – 1.3 (0.9 ± 0.1) | |||
| Man | 2.7 – 3.3 (3.0 ± 0.2) | 2.8 – 3.2 (3.0 ± 0.1) | 2.8 – 3.5 (3.0 ± 0.2) | |||
| Glycation | % Glycation at LC | 2.0 – 2.7 (2.3 ± 0.2) | 1.6 – 1.8 (1.7 ± 0.1) | 2.0 – 2.5 (2.2 ± 0.1) | ||
| % Glycation at HC | 3.3 – 4.8 (3.6 ± 0.4) | 2.6 – 3.0 (2.6 ± 0.2) | 3.1 – 4.4 (3.6 ± 0.4) | |||