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载脂蛋白B-48:使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、免疫印迹法和酶联免疫吸附测定法对血脂正常个体空腹浓度的比较

Apolipoprotein B-48: comparison of fasting concentrations measured in normolipidaemic individuals using SDS-PAGE, immunoblotting and ELISA.

作者信息

Jackson Kim G, Williams Christine M

机构信息

Hugh Sinclair Unit of Human Nutrition, School of Food Biosciences, The University of Reading, Reading, Berkshire, UK.

出版信息

Atherosclerosis. 2004 Oct;176(2):207-17. doi: 10.1016/j.atherosclerosis.2004.02.003.

Abstract

Raised levels of chylomicrons and chylomicron remnants, which circulate following a meal, have been implicated in the development of atherosclerosis. Apolipoprotein (apo) B-48 is exclusively associated with chylomicron particles and provides a specific direct measurement of the number of intestinally derived lipoproteins in the circulation. The quantification of apo B-48 in biological samples is difficult due to the very low concentration in plasma, structural similarity to the N-terminal 48% of apo B-100 and lack of an appropriate standard for apo B-48. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), followed by coomassie blue staining, has been used for many years to measure apo B-48 levels in triacylglycerol (TAG)-rich lipoprotein samples. The raising of antiserum to apo B-48 has led to development of more sensitive and specific methods including immunoblotting and enzyme-linked immunosorbant assays (ELISAs). This has enabled direct measurement of apo B-48 in plasma without the need for separation into TAG-rich lipoproteins. A high degree of variability was observed in the apo B-48 concentrations reported in the literature both within and between the SDS-PAGE, immunoblotting and ELISA methods.

摘要

进食后循环的乳糜微粒和乳糜微粒残粒水平升高与动脉粥样硬化的发展有关。载脂蛋白(apo)B-48仅与乳糜微粒颗粒相关,可直接特异性测量循环中肠道来源脂蛋白的数量。由于血浆中浓度极低、与apo B-100的N端48%结构相似且缺乏合适的apo B-48标准品,生物样品中apo B-48的定量分析很困难。多年来,十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)结合考马斯亮蓝染色用于测量富含三酰甘油(TAG)的脂蛋白样品中的apo B-48水平。针对apo B-48抗血清的产生促使了包括免疫印迹和酶联免疫吸附测定(ELISA)在内的更灵敏、特异方法的发展。这使得无需将血浆分离成富含TAG的脂蛋白就能直接测量血浆中的apo B-48。文献报道的SDS-PAGE、免疫印迹和ELISA方法中,apo B-48浓度在方法内部和方法之间均存在高度变异性。

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