权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Novel diagnosis of oxidized LDL in serum using monoclonal antibody

使用单克隆抗体诊断血清中氧化低密度脂蛋白的新方法

基本信息

批准号：
10557225
负责人：
TAKANO Tatsuya
金额：
$ 7.94万
依托单位：
Teikyo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2001
项目状态：
已结题

项目摘要

Our increasing knowledge of the atherogenic mechanisms of oxidation of LDL in the arterial wall, the clinical importance of circulating OxLDL is poorly understood, mainly because of a lack of a sensitive method to specifically detect circulating OxLDL levels. Although a number of investigators have measured thio-barbituric acid reactive substances in plasma, such as lipid hydroperoxides and isoprostanes, none have directly measured the formation of oxidatively modified LDL. Recently, immunoassays using murine monoclonal antibodies prepared against malondialdehyde-modified LDL (MDA-LDL) and against copper OxLDL demonstrated that circulating OxLDL or MDA-LDL exists in human plasma, and higher levels are found in patients with coronary heart disease (CHD). these studies, however, OxLDL was not distinguished from MDA-LDL because the monoclonal antibody against copper OxLDL cross-reacts against MDA-LDL. It is becoming clear that a large number of reactive lipid peroxidation products are generated in vivo during the oxidation of LDL. A study on apoE knockout mice, an animal model for the development of atherosclerosis, that auto-antibodies against OxLDL recognize different epitopes of complex structures formed during oxidation of lipoproteins or epitopes formed independently at different lesion sites.We have developed a unique monoclonal antibody FOHla/DLH3 that reacts specifically against oxidized phosphatidylcholine (OxPC) by use of a homogenate of atheromatous plaques of human aorta as an antigen but not against native LDL, MDA-LDL, acetylated LDL, or glycated LDL. By use of this antibody in combination with an anti-apoprotein B antibody, an enzyme immunoassay of circulating OxLDL was developed. The present study, the clinical characteristics of plasma OxLDL levels in patients with atherosclerotic disease were investigated.

我们对动脉壁中LDL氧化致动脉粥样硬化机制的了解越来越多，但循环OxLDL的临床重要性却知之甚少，主要是因为缺乏一种敏感的方法来特异性检测循环OxLDL水平。尽管许多研究者已经测量了血浆中的硫代巴比妥酸反应物质，如脂质氢过氧化物和异前列腺素，但没有人直接测量氧化修饰LDL的形成。最近，用抗丙二醛修饰低密度脂蛋白（MDA-LDL）和抗铜OxLDL的小鼠单克隆抗体制备的免疫分析表明，循环中的OxLDL或MDA-LDL存在于人血浆中，并且在冠心病（CHD）患者中含量较高。然而，这些研究并没有将OxLDL与MDA-LDL区分开来，因为抗铜单克隆抗体OxLDL与MDA-LDL发生交叉反应。越来越清楚的是，体内LDL氧化过程中会产生大量活性脂质过氧化产物。一项针对apoE敲除小鼠（动脉粥样硬化发展的动物模型）的研究表明，抗OxLDL的自身抗体识别脂蛋白氧化过程中形成的复杂结构的不同表位或在不同病变部位独立形成的表位。我们开发了一种独特的单克隆抗体FOHla/DLH3，通过使用人主动脉动脉粥样硬化斑块匀浆作为抗原，特异性对抗氧化磷脂酰胆碱（OxPC），但不对抗天然LDL， MDA-LDL，乙酰化LDL或糖化LDL。利用该抗体与抗载脂蛋白B抗体联合，建立了循环OxLDL的酶免疫测定方法。本研究旨在探讨动脉粥样硬化性疾病患者血浆OxLDL水平的临床特点。