Table 1.
Comparison of Identified HCPs Associated With mAbs 1–4
| Identified HCP | MW | pI | HCP Mass (ng/20 mg mAb) |
|||
|---|---|---|---|---|---|---|
| mAb1 | mAb2 | mAb3 | mAb4 | |||
| 14-3-3 protein epsilon | 7.89 | 6.52 | 9.0 | 24.3 | ND | ND |
| 40S ribosomal protein S15a | 14.83 | 10.13 | 9.0 | ND | ND | ND |
| 40S ribosomal protein S17 | 15.51 | 9.85 | ND | 20.3 | ND | 10.3 |
| 40S ribosomal protein S20 | 15.21 | 9.94 | ND | 20.3 | 2.3 | 8.7 |
| 40S ribosomal protein S21 | 9.14 | 8.32 | 9.0 | ND | ND | ND |
| 40S ribosomal protein SA | 19.72 | 9.29 | 18.0 | 28.4 | 2.3 | 10.9 |
| 60S acidic ribosomal protein P0* | 29.87 | 8.51 | 9.0 | 20.3 | 2.3 | 7.9 |
| 60S acidic ribosomal protein P2 | 11.67 | 4.54 | 24.2 | 26.3 | 3.0 | 13.5 |
| 60S ribosomal protein L12 | 14.69 | 9.55 | 12.6 | 20.3 | 2.3 | 10.3 |
| 60S ribosomal protein L23a | 23.11 | 10.45 | ND | ND | 2.3 | 10.3 |
| 78 kDa glucose-regulated protein†‡ | 72.33 | 5.16 | 9.0 | 26.3 | 3.0 | 10.3 |
| Acid trehalase-like protein 1 | 29.46 | 5.62 | ND | ND | ND | 9.5 |
| Actin, alpha cardiac muscle 1†‡ | 41.99 | 5.39 | 9.0 | 20.3 | 2.3 | 7.9 |
| actin, cytoplasmic 1†‡ | 41.71 | 5.39 | 13.4 | 20.3 | 3.3 | 7.9 |
| Alpha-enolase†‡ | 46.67 | 6.16 | 12.6 | 46.6 | 5.4 | 34.8 |
| Annexin A2 | 27.02 | 5.97 | 10.8 | ND | ND | ND |
| Aspartyl aminopeptidase | 51.91 | 7.12 | ND | ND | 2.3 | ND |
| Biglycan | 41.61 | 7.31 | 19.7 | 20.3 | 2.3 | 9.5 |
| Cathepsin B | 37.48 | 6.13 | 9.0 | 24.3 | 2.5 | 11.9 |
| Clusterin†‡ | 51.72 | 5.74 | 12.6 | 20.3 | 2.3 | 7.9 |
| Cofilin-1† | 18.52 | 8.09 | 22.4 | 24.3 | 3.6 | 11.1 |
| Cullin-associated NEDD8-dissociated protein 1 | 133.54 | 5.74 | 9.0 | 28.4 | 2.3 | 7.9 |
| Elongation factor 2† | 95.26 | 6.83 | 9.0 | 20.3 | 2.3 | 7.9 |
| Eukaryotic translation initiation factor 2 subunit 1 | 36.13 | 5.14 | ND | 22.3 | 3.8 | 17.4 |
| Galectin-3 | 32.42 | 7.37 | ND | ND | ND | 15.8 |
| Glutathione S-transferase P 1* | 23.62 | 7.80 | 9.0 | 26.3 | 4.5 | 15.8 |
| Glyceraldehyde-3-phosphate dehydrogenase† | 35.73 | 8.34 | 16.1 | 40.5 | 4.3 | 18.2 |
| GTP-binding nuclear protein Ran | 24.47 | 8.34 | 12.6 | 20.3 | 2.3 | ND |
| Guanine nucleotide-binding protein beta-2-like 1 | 30.44 | 7.39 | 9.0 | 20.3 | ND | ND |
| Heat shock cognate 71 kDa protein†,‡ | 70.76 | 5.36 | 9.0 | 22.3 | 2.3 | 7.9 |
| Histidine-tRNA ligase | 57.39 | 6.95 | ND | 20.3 | ND | ND |
| Histidyl-tRNA synthetase, cytoplasmic | 40.67 | 5.08 | ND | 20.3 | ND | ND |
| Histone H2A type 1 | 28.45 | 11.03 | 15.2 | ND | ND | ND |
| Histone H2A type 4 | 14.24 | 11.02 | ND | 30.4 | 2.3 | ND |
| Histone H2B type 1-N | 13.93 | 10.32 | 15.2 | ND | 2.3 | ND |
| Lactadherin | 16.14 | 9.41 | 28.7 | 34.4 | 3.2 | 14.3 |
| L-lactate dehydrogenase A chain | 36.50 | 7.42 | 9.0 | ND | ND | ND |
| Phosphotyrosine protein phosphatase | 17.90 | 6.01 | ND | 20.3 | 2.3 | 8.7 |
| Matrix metalloproteinase-19 | 58.90 | 7.88 | 9.0 | ND | ND | ND |
| Metalloproteinase inhibitor 1‡ | 22.39 | 8.47 | 9.0 | 20.3 | 2.3 | 7.9 |
| Nidogen-1†,‡ | 30.07 | 8.07 | 10.8 | ND | ND | ND |
| Nucleoside diphosphate kinase A | 17.18 | 6.33 | ND | ND | 7.9 | ND |
| Nucleoside diphosphate kinase B | 17.33 | 7.99 | ND | ND | 4.3 | ND |
| Out at first protein-like | 17.72 | 8.06 | 9.0 | 20.3 | 3.0 | 7.9 |
| Peptidyl-prolyl cis-trans isomerase A * | 17.89 | 8.28 | 9.0 | 20.3 | 2.3 | 7.9 |
| Peroxiredoxin-1†‡ | 22.25 | 8.05 | 27.8 | 46.6 | 4.3 | 19.8 |
| Phosphoglycerate mutase 1* | 20.19 | 8.12 | 9.8 | ND | ND | ND |
| Plectin-1 | 83.37 | 8.54 | ND | 40.5 | ND | ND |
| Proteasome subunit alpha type-7 | 23.76 | 8.48 | ND | ND | 2.3 | 7.9 |
| Protein disulfide-isomerase A6 | 28.38 | 4.91 | 9.0 | 46.6 | 3.8 | 15.8 |
| Protein S100-A6 | 10.00 | 5.48 | 9.0 | ND | ND | ND |
| Putative phospholipase B-like 2 | 65.50 | 6.28 | 9.0 | 36.5 | 3.0 | ND |
| Pyruvate kinase isozymes M1/M2†,‡ | 51.53 | 7.68 | 29.6 | 54.7 | 5.6 | 23.8 |
| Septin-2 | 37.05 | 6.18 | 9.0 | 20.3 | 2.3 | 7.9 |
| Serine protease HTRA1†,‡ | 28.70 | 7.03 | 9.0 | 20.3 | 2.3 | 7.9 |
| Serpin H1 | 46.53 | 8.68 | 9.0 | 20.3 | 2.3 | 7.9 |
| Sulfated glycoprotein 1†,‡ | 27.36 | 5.49 | 9.0 | 38.5 | 3.8 | 9.5 |
| T-complex protein 1 subunit alpha†,‡ | 60.30 | 5.99 | 14.4 | 24.3 | 2.5 | 9.5 |
| T-complex protein 1 subunit beta†,‡ | 57.45 | 6.46 | 16.1 | 36.5 | 3.6 | 15.8 |
| T-complex protein 1 subunit delta†,‡ | 42.11 | 8.27 | 9.8 | 20.3 | 2.3 | 7.9 |
| T-complex protein 1 subunit eta†,‡ | 54.87 | 7.42 | 9.8 | 20.3 | 2.3 | 7.9 |
| T-complex protein 1 subunit gamma†,‡ | 60.58 | 6.64 | 13.5 | 28.4 | 2.5 | 10.3 |
| T-complex protein 1 subunit theta† | 22.21 | 4.91 | 20.6 | 30.4 | 4.5 | 12.7 |
| T-complex protein 1 subunit zeta†,‡ | 57.94 | 6.90 | 11.7 | 20.3 | 2.5 | 7.9 |
| Triosephosphate isomerase | 16.42 | 8.19 | ND | 30.4 | 4.5 | 15.8 |
| Vimentin‡ | 44.50 | 4.81 | 18.0 | 24.3 | 2.3 | 10.3 |
| V-type proton ATPase subunit C 1‡ | 43.89 | 7.46 | ND | 20.3 | ND | ND |
Gray-shaded HCPs bound to all four mAbs. HCPs of potential risk to cause enzymatic degradation of the mAbs are indicated in bold text. ND indicates no detection of the HCPs or levels below 1% of the total HCP mass as calculated in Methods section.
*
HCPs previously found in Protein A-purified mAb product by Zhang et al.23
†
HCPs previously found in Protein A-purified mAb product by Doneanu et al.22
‡
HCPs associated with other mAbs identified in Levy et al.16