Methylglyoxal (MGO) produced during glycolysis is known to react with arginine residues on proteins to generate advanced glycation end products, such as N^δ-(5-hydro-5-methyl-4-imidazolone-2-yl)-ornithine (MG-H1). Since the production of MGO is increased during hyperglycemia or metabolic disorders in vivo, it is considered that the measurement of MG-H1 is useful for evaluating abnormalities in carbohydrate metabolism. Thus, we prepared a monoclonal antibody against MG-H1 to develop a conventional measurement system for MG-H1. Reactivity and specificity of the antibody to MGO-modified protein were confirmed by enzyme-linked immunosorbent assay and western blotting, respectively. The measurement of MG-H1 content by the antibody was positively correlated with that by electrospray ionization-liquid chromatography-tandem mass spectrometry and the ratio of modified arginine residues by amino acid analysis. Our results demonstrated that immunochemical methods could be useful for the estimation of MG-H1 content in modified proteins.

已知在糖酵解过程中产生的甲基乙二醛 (MGO) 与蛋白质上的精氨酸残基反应生成高级糖基化终产物，例如N ^δ -(5-hydro-5-methyl-4-imidazolone-2-yl)-鸟氨酸 (MG- H1)。由于在体内高血糖或代谢紊乱期间 MGO 的产生增加, 认为 MG-H1 的测量可用于评估碳水化合物代谢异常。因此，我们制备了针对 MG-H1 的单克隆抗体，以开发用于 MG-H1 的常规测量系统。分别通过酶联免疫吸附测定和蛋白质印迹证实了抗体对 MGO 修饰蛋白的反应性和特异性。抗体对MG-H1含量的测定与电喷雾-液相色谱-串联质谱法的测定值和氨基酸分析的修饰精氨酸残基比例呈正相关。我们的结果表明，免疫化学方法可用于估计修饰蛋白中的 MG-H1 含量。