title: Microheterogeneity of a purified IgG1 due to asymmetric Fab glycosylation.
abstract: A murine monoclonal anti-granulocyte IgG1, IMMU-MN3, was seen to exhibit heterogeneity. On reduced SDS-PAGE, the purified antibody appeared as two heavy-chain bands of unequal intensity, and only one light-chain band. Hydrophobic interaction chromatography (HIC) also resolved two populations of the IMMU-MN3 antibody. Based on Concanavalin A affinity chromatography, enzymatic digestion with Endoglycosidase F and carbohydrate analysis, it was found that the heterogeneity detected by SDS-PAGE and HIC was due to differences in glycosylation. Furthermore, sequential gel analysis (non-reduced/reduced) demonstrated that the upper heavy-chain band was asymmetrically glycosylated.
authors.affiliation: Immunomedics Inc., Newark, NJ 07103.
authors.forename: R C
authors.initials: RC
authors.lastname: Grebenau
authors.forename: D M
authors.initials: DM
authors.lastname: Goldenberg
authors.forename: C H
authors.initials: CH
authors.lastname: Chang
authors.forename: G A
authors.initials: GA
authors.lastname: Koch
authors.forename: D V
authors.initials: DV
authors.lastname: Gold
authors.forename: A
authors.initials: A
authors.lastname: Kunz
authors.forename: H J
authors.initials: HJ
authors.lastname: Hansen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry*; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000494:pharmacology; D051379:Mice
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D046911:Macromolecular Substances; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase