title: Usefulness of antibodies for evaluating the biological significance of AGEs.
abstract: Polyclonal and monoclonal antibodies have been widely applied to demonstrate the presence of advanced glycation end products (AGEs) in vivo. However, our previous study showed that monoclonal anti-AGE antibody (6D12) and polyclonal anti-N epsilon-(carboxymethyl)lysine (CML) antibody recognize not only CML but also N epsilon-(carboxyethyl)lysine (CEL), thus indicating that we should pay attention to the specificity of the antibodies. As a result, we prepared specific monoclonal antibodies against CML, CEL, N omega-(carboxymethyl)arginine (CMA), and S-(carboxymethyl)cysteine (CMC). Our immunochemical study using anti-CMA antibody demonstrated that the CMA content increased in a time-dependent manner when collagen was incubated with glucose, indicating that immunological quantification using the specific antibody is especially useful for measuring an acid-labile AGE structure, such as CMA. Monoclonal antibody is also applied to identify a novel biological marker in pathological lesions. We prepared antibody libraries against proteins modified with aldehydes, such as glyoxal, methylglyoxal, and glycolaldehyde (GA), and one antibody, GA5, which specifically reacts with the GA-modified protein that is recognized in human atherosclerotic lesions. Following successive high-performance liquid chromatography purification, the GA5-reactive compound was isolated and its chemical structure was found to be 3-hydroxy-4-hydroxymethyl-1-(5-amino-5-carboxypentyl) pyridinium cation, which was named GA-pyridine. Taken together, these results demonstrate that a specific antibody is a powerful tool for analyzing novel biomarkers, formation pathways, and the efficacy of AGE inhibitors.
authors: Department of Medical Biochemistry, Faculty of Medical and Pharmaceutical Sciences, Kumamoto University, Kumamoto 860-8556, Japan. nagai-883@umin.ac.jp
authors: Ryoji
authors: R
authors: Nagai
authors: Yukio
authors: Y
authors: Fujiwara
authors: Katsumi
authors: K
authors: Mera
authors: Keita
authors: K
authors: Motomura
authors: Yasunori
authors: Y
authors: Iwao
authors: Keiichiro
authors: K
authors: Tsurushima
authors: Mime
authors: M
authors: Nagai
authors: Kazuhiro
authors: K
authors: Takeo
authors: Makiko
authors: M
authors: Yoshitomi
authors: Masaki
authors: M
authors: Otagiri
authors: Tsuyoshi
authors: T
authors: Ikeda
mesh_terms: D000447:Aldehydes / Q000032:analysis / Q000276:immunology; D000906:Antibodies; D001120:Arginine / Q000031:analogs & derivatives / Q000032:analysis; D017127:Glycation End Products, Advanced / Q000032:analysis* / Q000276:immunology; D008239:Lysine / Q000031:analogs & derivatives / Q000032:analysis
chemical_list: D000447:Aldehydes; D000906:Antibodies; D017127:Glycation End Products, Advanced; C408696:N(omega)-carboxymethylarginine; C048496:N(6)-carboxymethyllysine; D001120:Arginine; D008239:Lysine