Sensitive detection of oxidatively modified low density lipoprotein using a monoclonal antibody.
Sensitive detection of oxidatively modified low density lipoprotein using a monoclonal antibody.
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发表时间：
1996
影响因子：
6.5
通讯作者：
H. Itabe;Hisashi Yamamoto;T. Imanaka;Kyoko Shhamura;H. Uchiyama;J. Kimura;T. Sanaka;Yoshiya Hat;T. Takano
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作者：
H. Itabe;Hisashi Yamamoto;T. Imanaka;Kyoko Shhamura;H. Uchiyama;J. Kimura;T. Sanaka;Yoshiya Hat;T. Takano
我们建立了一种能够测定极低浓度氧化低密度脂蛋白（OxLDL）的新方法。在我们以前的研究中，我们获得了一种新的抗氧化脂蛋白的鼠单克隆抗体（Itabe，H.等，1994年。J.Biol.Chem.269：15274-15279）。该抗体的表位存在于磷脂酰胆碱的氧化产物中，其可与多肽（包括载脂蛋白B）形成复合物。当在使用抗人载脂蛋白B抗体进行夹心ELISA之前将单克隆抗体预包被到微量滴定威尔斯孔上时，可以检测到0.5 ng铜诱导的OxLDL蛋白。OxLDL的检测依赖于单克隆抗体的存在，并被氧化磷脂酰胆碱（OxPC）阻断。在相同的夹心ELISA条件下，天然LDL表现出剂量依赖性的吸光度增加，OxPC与BSA的复合物抑制。这些结果表明，可能发生的氧化修饰的人血浆LDL，这是公认的抗OxPC抗体。发现正常人受试者的LDL氧化水平为0.52 ± 0.35单位/5 μ g LDL蛋白，其中1单位定义为通过该测定对应于1 ng铜诱导的OxLDL的反应性。此外，我们还发现，接受血液透析治疗的患者的LDL氧化水平比正常人高出8倍以上。提示人血浆中的LDL在一定条件下发生氧化修饰，这种测定OxLDL的方法可用于研究体内氧化反应与各种病理状态的关系。
We have established a new method capable of measuring the very low concentrations of oxidized low density lipoprotein (OxLDL). In our previous study, we obtained a novel murine monoclonal antibody against oxidized lipoproteins (Itabe, H. et al. 1994. J. Biol. Chem. 269: 15274-15279). The epitope of this antibody resides in oxidized products of phosphatidylcholine that can form complexes with polypeptides, including apolipoprotein B. When the monoclonal antibody was precoated onto microtiter wells prior to carrying out a sandwich ELISA using an anti-human apolipoprotein B antibody, it was possible to detect 0.5 ng protein of copper-induced OxLDL. The detection of OxLDL was dependent on the presence of monoclonal antibody and was blocked by oxidized phosphatidylcholine (OxPC). Under the same sandwich ELISA condition, native LDL showed a dose-dependent increase of absorbance that was inhibited by complex of OxPC with BSA. These results suggest the possible occurrence of oxidative modification of human plasma LDL, which is recognized by the antibody against OxPC. The level of LDL oxidation of normal human subjects was found to be 0.52 +/- 0.35 units per 5 mu g protein of LDL, where one unit was defined as the reactivity corresponding to 1 ng of copper-induced OxLDL by this assay. Furthermore, we found that the LDL oxidation level in patients who had been receiving hemodialysis treatment was increased more than eightfold over that of normal subjects. We suggest that LDL in human plasma is oxidatively modified under certain conditions and this method for measurement of OxLDL could be used to study the relationship between in vivo oxidation reaction and various pathological conditions.