title: A global comparability approach for biosimilar monoclonal antibodies using LC-tandem MS based proteomics.
abstract: Liquid chromatography-tandem mass spectrometry-based proteomics for peptide mapping and sequencing was used to characterize the marketed monoclonal antibody trastuzumab and compare it with two biosimilar products, mAb A containing D359E and L361M variations at the Fc site and mAb B without variants. Complete sequence coverage (100%) including disulfide linkages, glycosylations and other commonly occurring modifications (i.e., deamidation, oxidation, dehydration and K-clipping) were identified using maps generated from multi-enzyme digestions. In addition to the targeted comparison for the relative populations of targeted modification forms, a non-targeted approach was used to globally compare ion intensities in tryptic maps. The non-targeted comparison provided an extra-dimensional view to examine any possible differences related to variants or modifications. A peptide containing the two variants in mAb A, D359E and L361M, was revealed using the non-targeted comparison of the tryptic maps. In contrast, no significant differences were observed when trastuzumab was self-compared or compared with mAb B. These results were consistent with the data derived from peptide sequencing via collision induced dissociation/electron transfer dissociation. Thus, combined targeted and non-targeted approaches using powerful mass spectrometry-based proteomic tools hold great promise for the structural characterization of biosimilar products.
authors.affiliation: Department of Chemistry, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Shun-Li
authors.initials: SL
authors.lastname: Chen
authors.forename: Shiaw-Lin
authors.initials: SL
authors.lastname: Wu
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Huang
authors.forename: Jia-Bao
authors.initials: JB
authors.lastname: Huang
authors.forename: Shu-Hui
authors.initials: SH
authors.lastname: Chen
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000032:analysis*; D059451:Biosimilar Pharmaceuticals / Q000032:analysis*; D016466:CHO Cells; D002853:Chromatography, Liquid / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D010449:Peptide Mapping / Q000379:methods; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods; D000068878:Trastuzumab
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D059451:Biosimilar Pharmaceuticals; D000068878:Trastuzumab