title: Therapeutic antibody glycosylation analysis: a contract research organization perspective in the frame of batch release or comparability support.
abstract: Glycosylation of the Fc moiety of a monoclonal antibody is a heterogeneous posttranslational process considered as a critical quality attribute of the purified drug substance due to its major impact on safety and efficacy (i.e., immunogenicity, CDC or ADCC effector functions, etc.). Glycosylation should thus be addressed for batch-to-batch comparability and for drug substance characterization, in terms of identity and/or purity testing. We present below a set of efficient, performing and complementary analytical tests that can be used alone or in combination, depending on the information needed and available laboratory instrumentation. The results obtained using these techniques for "global" glycosylation profile, N-glycans profiling, monosaccharides, and sialic acids determination are presented for the Trastuzumab (Herceptin)-humanized mAb produced in CHO.
authors.affiliation: Quality Assistance, Donstiennes, Belgium.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Delobel
authors.forename: Fabrice
authors.initials: F
authors.lastname: Cantais
authors.forename: Anicet
authors.initials: A
authors.lastname: Catrain
authors.forename: Erell
authors.initials: E
authors.lastname: Dereux
authors.forename: Géry
authors.initials: G
authors.lastname: Van Vyncht
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D005947:Glucose / Q000737:chemistry / Q000302:isolation & purification; D006020:Glycopeptides / Q000737:chemistry* / Q000302:isolation & purification; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D012015:Reference Standards; D012794:Sialic Acids / Q000737:chemistry / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods / Q000592:standards; D013194:Staining and Labeling; D000068878:Trastuzumab
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D006020:Glycopeptides; D011134:Polysaccharides; D012794:Sialic Acids; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D005947:Glucose; D000068878:Trastuzumab