Waeg G, Dimsity G, Esterbauer H
Institute of Biochemistry, University of Graz, Austria.
Free Radic Res. 1996 Aug;25(2):149-59. doi: 10.3109/10715769609149920.
A promising approach to study lipid peroxidation pathology is antibodies recognizing aldehydes which react with and became bound to amino acid side chains of proteins. We present in this study the characterization of several monoclonal antibodies which recognize 4-hydroxynonenal (HNE) modified proteins. Six out of 20 antibodies recognizing HNE modified BSA were able to detect HNE-protein adducts in peroxidized liver microsomes. Two of these antibodies were selected and characterized. Both antibodies could also detect HNE-protein adducts in oxidized low density lipoprotein. They exhibit no detectable cross reaction with proteins modified by malonaldehyde, nonanal, nonanal and 4-hydroxyhexenal. Protein bound 4-hydroxyoctenal and 4-hydroxydecenal were recognized to some extent. Further characterization revealed that the two antibodies are highly selective for HNE bound to histidine with only some cross reaction to HNE bound to lysine and cysteine. Preliminary quantitative ELISA-analysis showed that oxidized microsomes and oxidized LDL contain 12 nmol and 3 nmol HNE-histidine per mg protein respectively.
一种研究脂质过氧化病理的有前景的方法是使用能识别醛类的抗体，这些醛类会与蛋白质的氨基酸侧链发生反应并与之结合。在本研究中，我们对几种识别4-羟基壬烯醛（HNE）修饰蛋白的单克隆抗体进行了表征。在识别HNE修饰牛血清白蛋白（BSA）的20种抗体中，有6种能够检测过氧化肝微粒体中的HNE-蛋白加合物。我们选择并表征了其中两种抗体。这两种抗体还能检测氧化型低密度脂蛋白中的HNE-蛋白加合物。它们与丙二醛、壬醛、壬醛和4-羟基己烯醛修饰的蛋白未表现出可检测到的交叉反应。蛋白结合的4-羟基辛烯醛和4-羟基癸烯醛在一定程度上能被识别。进一步的表征表明，这两种抗体对与组氨酸结合的HNE具有高度选择性，对与赖氨酸和半胱氨酸结合的HNE只有一些交叉反应。初步的定量酶联免疫吸附测定（ELISA）分析表明，氧化微粒体和氧化型低密度脂蛋白分别含有每毫克蛋白12纳摩尔和3纳摩尔的HNE-组氨酸。
