Table II.
Analytical Similarity Assessment between ABP 710 and Infliximab
| Analytical Testing/Attributes | ABP 710 (n) | Infliximab US (n) | Infliximab EU (n) |
|---|---|---|---|
| Intact molecular weight (ppma) | |||
| Peak A: A1G0F:A2G0F, 148,310 Da | 5 (1) | 5 (1) | 0 (1) |
| Peak B: A2G0F:A2G0F, 148,514 Da | 2 (1) | 11 (1) | 6 (1) |
| Peak C. A2G0F:A2G0F + 1 K, 148642 Da | 3 (1) | 21 (1) | 20 (1) |
| Peak D: A2G0F:A2G1F, 148,676 Da | 5 (1) | 12 (1) | 13 (1) |
| Peak E: A2G0F:A2G0F + 2 K, 148770 Da | 10 (1) | 8 (1) | 5 (1) |
| Peak F: A2G0F:A2G1F + 1 K, 148804 Da | 10 (1) | 16 (1) | 18 (1) |
| Peak G: A2G1F:A2G1F or A2G0F:A2G2F, 148,838 Da | 7 (1) | 29 (1) | 20 (1) |
| Peak H: A2G0F:A2G1F + 2 K, 148932 Da | 9 (1) | 8 (1) | 2 (1) |
| Peak I: A2G1F:A2G1F or A2G0F:A2G2F + 2 K, 149094 Da | 0 (1) | 10 (1) | 3 (1) |
| Reduced and deglycosylated HC, 49389 Da | 4 (1) | 17 (1) | 22 (1) |
| Reduced and deglycosylated HC + K, 49517 Da | 4 (1) | 24 (1) | 18 (1) |
| Reduced and deglycosylated LC, 23439 Da | 3 (1) | 26 (1) | 27 (1) |
| Glycan Map (%) | |||
| High Mannose | 3.2–4.2 (14) | 3.0–5.5 (28) | 3.1–5.5 (20) |
| Afucosylation | 6.1–7.3 (14) | 6.7–11.1 (28) | 6.9–10.9 (20) |
| β-galactosylation | 25.1–34.2 (14) | 25.9–52.0 (28) | 25.7–51.2 (20) |
| α-galactosylation | N/A | 1.6–6.6 (28) | 1.5–6.1 (20) |
| Sialylation | 0.9–1.6 (14) | 3.1–10.0 (28) | 4.1–9.7 (20) |
| Isoelectric point | 7.4 (1) | 7.4 (1) | 7.4 (1) |
| FTIR/spectral similarity (%) | |||
| US RP | 99.7–99.8 (6) | 99.7–100 (3) | 99.7–99.9 (3) |
| EU RP | 99.6–99.9 (6) | 99.7–99.9 (3) | 99.6–100 (3) |
| NUV-CD/spectral similarity (%) | |||
| US RP | 97.5–99.1 (6) | 98.9–100 (3) | 98.7–99.2 (3) |
| EU RP | 97.1–99.3 (6) | 98.2–98.7(3) | 98.3–100 (3) |
| DSC (°C) | |||
| Tm1 | 67.7–67.9 (9) | 67.6–67.8 (6) | 67.6–67.8 (6) |
| Tm2 | 83.3–84.3 (9) | 83.3–84.2 (6) | 83.2–84.3 (6) |
| Size Variants by SE-UHPLC (%) | |||
| Main Peak | 98.6–99.2 (12) | 99.3–99.8 (28) | 99.5–99.8 (19) |
| Size Variants by rCE-SDS | |||
| Heavy Chain + Light Chain | 96.6–97.3 (12) | 98.2–98.9 (28) | 98.4–98.9 (19) |
| Fragments | 0.7–1.0 (12) | 0.3–1.1 (28) | 0.4–0.8 (19) |
| Non-glycosylated heavy chain | 1.6–2.1 (12) | 0.4–0.6 (28) | 0.4–0.6 (19) |
| Size Variants by nrCE-SDS | |||
| Main Peak | 97.1–98.1 (12) | 97.8–98.9 (28) | 98.4–98.9 (12) |
| Charge Variants by CEX-UHPLC | |||
| Main Peak | 43.5–47.7 (12) | 31.7–51.7 (28) | 37.2–45.4(18) |
| Acidic Peaks | 19.5–23.6 (12) | 9.9–15.8 (28) | 9.6–17.7 (18) |
| Basic Peaks | 30.0–36.1 (12) | 34.2–56.2 (28) | 40.5–53.2 (18) |
| HIAC subvisible particle (particles/mL) | |||
| ≥ 2 μm | 40–569 (19) | 4568–7064 (7) | 3782–14,353 (7) |
| ≥ 5 μm | 5–138 (19) | 724–1341 (7) | 675–3138 (7) |
| ≥ 10 μm | 0–39 (19) | 64–172 (7) | 82–315 (7) |
| ≥ 25 μm | 0–2 (19) | 0–3 (7) | 1–3 (7) |
| MFI subvisible particle (particles/mL) | |||
| ≥ 5 μm particles | 19–1260 (19) | 952–3263 (7) | 758–6047 (7) |
| ≥ 5 μm non-spherical particles | 12–260 (19) | 328–1940 (7) | 384–3425 (7) |
| SV-AUC/monomer (%) | 98.4–99.4 (3) | 98.8–99.7 (3) | 99.1–99.9 (3) |
| SE-HPLC-LS MW (kDa) | |||
| Monomer | 150–151 (3) | 151–152 (3) | 151–152 (3) |
| Dimer | 284–287 (3) | 292–319 (3) | 285–334 (3) |
| Reconstituted protein concentration (mg/mL) | 9.1–9.6 (19) | 9.2–9.8 (26) | 9.1–9.7 (18) |
| Protein content (mg) | 96–99 (19) | 97–102 (26) | 96–101 (18) |
n = number of lots tested
appm = 1,000,000 x |(observed mass - theoretical mass)|/theoretical mass
Glycosylation status of observed monoclonal antibody species: A = number of antenna (GlcNAc) on trimannosyl core; F = Fucose; G = linked galactose on antenna; HC heavy chain, K = C-terminal lysine. LC light chain
Reconstituted protein concentration and protein content determined from 100 mg vial
SV-AUC sedimentation velocity analytical ultracentrifugation, CEX-UHPLC cation exchange ultra-high performance liquid chromatography, DSC differential scan-ning calorimetry, FTIR Fourier-transform infrared spectroscopy, HC heavy chain, LC light chain, MFI micro-flow imaging, NUV-CD near-UV circular dichroism, nrCE-SDS non-reduced capillary electrophoresis–sodium dodecyl sulfate, NGHC non-glycosylated heavy chain, rCE-SDS reduced capillary electrophoresis–sodium dodecyl sulfate, RP reference product, SE-UHPLC size exclusion untra high performance liquid chromatography, SE-HPLC-LS size exclusion high performance liquid chromatography with light scattering