Uchida K, Osawa T, Hiai H, Toyokuni S
Laboratory of Food and Biodynamics, Nagoya University School of Agriculture, Japan.
Biochem Biophys Res Commun. 1995 Jul 26;212(3):1068-73. doi: 10.1006/bbrc.1995.2078.
Among the aldehydes that originate from the peroxidation of cellular membrane lipids, 4-hydroxy-2-nonenal (HNE) is thought to be largely responsible for cytopathological effects observed during oxidative stress. Taking advantage of the fact that HNE is very reactive with proteins and forms stable Michael addition-type adducts, a novel immunochemical procedure for quantifying "free" HNE has been developed. The method designated as "HNE-trapping ELISA" is based on the detection of HNE trapped by a protein that has been coated in the immunoplate. The HNE-derived epitopes generated in the coating protein are then detected by the ELISA using a monoclonal antibody (mAbHNEJ-2) specific to the haptenic groups of the HNE-protein conjugates. Using this method, we determined that a considerable amount of HNE was released from human plasma low density lipoproteins (LDL) treated with copper ions or endothelial cells.
在源于细胞膜脂质过氧化的醛类中，4-羟基-2-壬烯醛（HNE）被认为在很大程度上是氧化应激期间观察到的细胞病理学效应的原因。利用HNE与蛋白质反应非常活跃并形成稳定的迈克尔加成型加合物这一事实，已开发出一种用于定量“游离”HNE的新型免疫化学方法。指定为“HNE捕获ELISA”的方法基于检测被包被在免疫板中的蛋白质捕获的HNE。然后使用对HNE-蛋白质缀合物的半抗原基团具有特异性的单克隆抗体（mAbHNEJ-2）通过ELISA检测包被蛋白中产生的HNE衍生表位。使用这种方法，我们确定在用铜离子处理的人血浆低密度脂蛋白（LDL）或内皮细胞中释放出大量的HNE。
