title: Effect of posttranslational modifications on the thermal stability of a recombinant monoclonal antibody.
abstract: The effect of oligosaccharides and C-terminal lysine residues on the thermal stability of a recombinant IgG(1) monoclonal antibody was investigated using differential scanning calorimetry (DSC). The C-terminal lysine did not appear to affect the thermal stability of this IgG(1) molecule. However, oligosaccharides, which are buried between the two CH(2) domains, provided significant stabilizing energy. Characterization of the Fab and Fc after papain digestion suggested that the stabilizing effect of oligosaccharides on this molecule was through stabilizing CH(2) domains. Oligosaccharides had little effect on the thermal stability of Fab region and CH(3) domains. It was also interesting to note that both intact IgG(1) antibody and its Fab, but not the Fc regions, appeared to form precipitate after thermal unfolding under the experimental conditions.
authors.affiliation: Process Sciences Department, Abbott Bioresearch Center, 100 Research Drive, Worcester, MA 01605, United States. hongcheng.liu@abbott.com <hongcheng.liu@abbott.com>
authors.forename: Hongcheng
authors.initials: H
authors.lastname: Liu
authors.forename: Georgeen-Gaza
authors.initials: GG
authors.lastname: Bulseco
authors.forename: Joanne
authors.initials: J
authors.lastname: Sun
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning / Q000379:methods; D006031:Glycosylation; D006358:Hot Temperature; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry*; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry; D010206:Papain / Q000737:chemistry; D011489:Protein Denaturation; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D009844:Oligosaccharides; D011994:Recombinant Proteins; D010206:Papain