MODIFICATION OF THE BICINCHONINIC ACID PROTEIN ASSAY TO ELIMINATE LIPID INTERFERENCE IN DETERMINING LIPOPROTEIN PROTEIN-CONTENT
MODIFICATION OF THE BICINCHONINIC ACID PROTEIN ASSAY TO ELIMINATE LIPID INTERFERENCE IN DETERMINING LIPOPROTEIN PROTEIN-CONTENT
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DOI：
10.1016/0003-2697(92)90248-6
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发表时间：
1992-08-01
影响因子：
2.9
通讯作者：
EVANS, TA
中科院分区：
文献类型：
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作者：
MORTON, RE;EVANS, TA
研究了二喹啉甲酸（BCA）测定蛋白质的方法，以确定其在测定血浆脂蛋白中蛋白质含量的实用性。尽管其它方法，主要是基于Lowry等（1951，J.Biol.Chem.193，265-275）的方法已被广泛用于此目的，但BCA方法对许多常见去污剂和缓冲剂的耐受性提供了优于基于Lowry的方法的明确优势。在本研究中，将通过BCA方法获得的脂蛋白值与Lowry等人程序的标准修改进行了比较，因为该试验构成了许多相关文献的基础。发现标准BCA测定法高估极低密度脂蛋白的蛋白质含量约70%，高估低密度脂蛋白的蛋白质含量约30%;高密度脂蛋白值相比有利。通过BCA测定法的过度估计可确定脂蛋白组分的相对磷脂含量。通过在分析前向样品中加入2%十二烷基硫酸钠消除了这种明显的脂质效应。在存在该洗涤剂的情况下，这三种脂蛋白组分的BCA测定值分别为参比测定值的97、90和98%。
The bicinchoninic acid (BCA) assay method for the determination of protein has been investigated for its utility in measuring the protein content of plasma lipoproteins. Although other methods, principally those based on the method of Lowry et al. (1951, J. Biol. Chem. 193, 265–275) have been extensively used for this purpose, the tolerance of the BCA method to many commonly encountered detergents and buffers offers a definite advantage over the Lowry-based methods. In this study, lipoprotein protein values obtained by the BCA method were compared to a standard modification of the Lowry et al. procedure since this assay forms the basis of much of the relevant literature. The standard BCA assay was found to overestimate the protein content of very low density lipoprotein by ∼70% and low density lipoprotein by ∼30%; high density lipoprotein values compared favorably. Overestimations by the BCA assay paralleled the relative phospholipid content of the lipoprotein fractions. This apparent lipid effect was eliminated by the addition of 2% sodium dodecyl sulfate to samples prior to the analysis. In the presence of this detergent, BCA assay measurements for these three lipoprotein fractions were 97, 90, and 98%, respectively, of the reference assay values.