RADIOIMMUNOASSAY STUDIES OF HUMAN APOLIPOPROTEIN-E
RADIOIMMUNOASSAY STUDIES OF HUMAN APOLIPOPROTEIN-E
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DOI：
10.1172/jci109975
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发表时间：
1980-01-01
影响因子：
15.9
通讯作者：
SCIACCA, R
中科院分区：
文献类型：
--
作者：
BLUM, CB;ARON, L;SCIACCA, R
本文报道了一种灵敏、特异的人载脂蛋白E（apoE）双抗体放射免疫分析法的建立和首次应用。采用肝素-琼脂糖亲和层析、DEAE-纤维素层析和制备型聚丙烯酰胺凝胶电泳从高脂血症患者的极低密度脂蛋白中纯化ApoE。纯化的载脂蛋白具有载脂蛋白E特征的氨基酸组成，当注射到兔子体内时会产生单特异性抗血清。在5 mM十二烷基硫酸钠存在下进行的放射免疫测定的工作范围为0.8-12 ng。试验内变异系数[CV]为9%，系统试验间变异性的CV为3%。先前用有机溶剂脱脂样品未改变其免疫反应性。在26名正常志愿者中，平均血浆apoE浓度为36 ± 0.01。13 μ g/ml。高脂血症患者（n = 68）的平均apoE水平较高。患有III型高脂蛋白血症的单个患者的血浆apoE水平为664 μ g/ml。在108例正常和非乳糜微粒血症高脂血症患者中，血浆apoE水平与血浆胆固醇和甘油三酯水平独立相关。这种关系的多重相关系数为0.73。因此，血浆胆固醇和甘油三酯浓度的变化描述了该人群中apoE浓度变化的53%。用琼脂糖柱层析和血浆超离心法研究载脂蛋白E在脂蛋白中的分布。琼脂糖柱层析证明，所有或几乎所有的血浆apoE与脂蛋白。在正常志愿者和高胆固醇血症患者的血浆中，发现apoE存在于2种离散的脂蛋白类别中：极低密度脂蛋白和一组大小和密度特征与HDL 2相似的脂蛋白颗粒。在高脂血症患者中，几乎所有的apoE都与富含脂蛋白的脂蛋白有关。
The development and 1st applications of a sensitive and specific double antibody radioimmunoassay for human apolipoprotein E (apoE) are described. ApoE was purified from the very low density lipoproteins of hypertriglyceridemic patients by heparin-agarose affinity chromatography, DEAE-cellulose chromatography and preparative polyacrylamide gel electrophoresis. The purified apoprotein had an amino acid composition characteristic of apoE and resulted in the production of monospecific antisera when injected into rabbits. The radioimmunoassay, which was carried out in the presence of 5 mM sodium dodecyl sulfate, had a working range of 0.8-12 ng. The within-assay coefficient of variation [CV] was 9% and the CV for systematic between-assay variability was 3%. Prior delipidation of samples with organic solvents did not alter their immunoreactivity. In 26 normal volunteers, the mean plasma apoE concentration was 36 .+-. 13 .mu.g/ml. Hyperlipidemic patients (n = 68) had higher mean apoE levels. A single patient with type III hyperlipoproteinemia had a plasma apoE level of 664 .mu.g/ml. The plasma apoE level was independently related to plasma cholesterol and triglyceride levels in a population of 108 normal and nonchylomicronemic hyperlipidemic patients. The multiple correlation coefficient for this relationship was 0.73. Thus, variation in plasma cholesterol and triglyceride concentrations described 53% of the variation in apoE concentrations in this population. The lipoprotein distribution of apoE was investigated by agarose column chromatography and ultracentrifugation of plasma. Agarose column chromatography demonstrated that all or nearly all plasma apoE is associated with lipoproteins. In plasma from normal volunteers and hypercholesterolemic patients, apoE was found in 2 discrete lipoprotein classes: very low density lipoproteins and a set of lipoprotein particles with size and density characteristics similar to HDL2. In hypertriglyceridemic patients, nearly all apoE was associated with the triglyceride-rich lipoproteins.