title: Characterization of antibodies to advanced glycosylation end products on protein.
abstract: Antibodies directed against advanced glycosylation end products (AGEs) formed during a Maillard reaction have been generated and characterized. Since protein-bound AGEs recognized by the antibodies were labile to acid hydrolysis, the antibodies were further characterized by using the AGE-alpha-acetyl-L-lysine methyl ester (AGE-ALME) with a brown and fluorescent property as well as the AGE-proteins. The antibodies reacted with fluorescent compounds, rather than brown pigment compounds, in the AGE-ALME. The fluorescent compounds in the AGE-ALME were separated into four fluorescent compounds by reversed-phase thin layer chromatography (TLC). Of the fluorescent compounds tested, compound 3 (Rf = 0.63), as designated on a TLC plate, showed the highest affinity for the antibodies. In addition, the antibody recognition to the cross-linked oligomers with fluorescence in the AGE-protein was investigated by using bovine pancreatic ribonuclease A (RNase), which is known as a model protein for studying AGE-induced cross-linking. Fluorescence in the AGE-RNase existed in both of the oligomers and the monomer. The cross-linked oligomers exhibited higher affinity to the antibodies than did the monomer, which has a similar degree of fluorescent intensity. These results indicate that our antibodies against cross-linked protein-bound AGEs may serve as a useful tool to elucidate pathophysiological roles of advanced Maillard reaction in diabetic complications and aging processes.
authors.affiliation: 2nd Department of Internal Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Nakayama
authors.forename: S
authors.initials: S
authors.lastname: Taneda
authors.forename: T
authors.initials: T
authors.lastname: Mitsuhashi
authors.forename: S
authors.initials: S
authors.lastname: Kuwajima
authors.forename: S
authors.initials: S
authors.lastname: Aoki
authors.forename: Y
authors.initials: Y
authors.lastname: Kuroda
authors.forename: K
authors.initials: K
authors.lastname: Misawa
authors.forename: K
authors.initials: K
authors.lastname: Yanagisawa
authors.forename: S
authors.initials: S
authors.lastname: Nakagawa
mesh_terms: D000906:Antibodies / Q000032:analysis*; D002850:Chromatography, Gel; D002855:Chromatography, Thin Layer; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D008239:Lysine / Q000031:analogs & derivatives* / Q000276:immunology; D015416:Maillard Reaction
chemical_list: D000906:Antibodies; C069694:alpha-acetyllysine methyl ester; D008239:Lysine