Development of a homogeneous assay to measure remnant lipoprotein cholesterol
Development of a homogeneous assay to measure remnant lipoprotein cholesterol
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DOI：
10.1373/clinchem.2007.092296
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发表时间：
2007-12-01
影响因子：
9.3
通讯作者：
Yamamura, Taku
中科院分区：
文献类型：
--
作者：
Miyauchi, Kazuhito;Kayahara, Norihiko;Yamamura, Taku
背景资料：富含甘油三酯脂蛋白（TRL）残留物的定量可用于冠状动脉疾病的风险评估和III型高脂蛋白血症的诊断。尽管近年来已广泛评价了残余物样颗粒胆固醇的免疫分离程序，但用于测量TRL残余物的可用方法尚未在常规实验室实践中获得广泛使用，这表明需要一种无需预处理即可测量血清或血浆中TRL残余胆固醇的均相测定法。我们筛选合适的表面活性剂，表现出良好的选择性对极低密度脂蛋白残基（VLDLR）馏分，包括中密度脂蛋白（IDL）。我们通过脂蛋白的凝胶过滤及其粒度分布研究了该测定法的主要特征。结果：聚氧乙烯-聚氧丁烯嵌段共聚物（POE-POB）对VLDLR和IDL组分具有良好的选择性。POE-POB在胆固醇酯酶（CHER）存在下从IDL颗粒中去除载脂蛋白（apo）E和apo C-III，反应后IDL的粒径分布变小。这些结果表明，IDL颗粒在CHER和POE-POB的存在下被特异性修饰，使得它们的组分胆固醇可用于酶促测定。磷脂酶D的加入提高了对乳糜微粒残留物（CMR）的反应性。结论：该方法可用于TRL残基胆固醇（CMRs、VLDLRs和IDLs）的测定，具有较高的灵敏度和特异性。(C)2007年美国临床化学协会。
Background: Quantification of triglyceride-rich lipoprotein (TRL) remnants is useful for risk assessment of coronary artery disease and the diagnosis of type III hyperlipoproteinemia. Although an immunoseparation procedure for remnant-like particle cholesterol has been evaluated extensively in recent years, available methods for measuring TRL remnants have not achieved wide use in routine laboratory practice, suggesting a need for a homogeneous assay that can measure TRL remnant cholesterol in serum or plasma without pretreatment.Methods: We screened for suitable surfactants that exhibited favorable selectivity toward the VLDL remnant (VLDLR) fraction, including intermediate-density lipoproteins (IDLs). We investigated the principal characteristics of this assay by gel filtration of lipoproteins and their particle size distribution. We developed a simple assay and evaluated its performance with the Hitachi-7170 analyzer.Results: Polyoxyethylene-polyoxybutylene block copolymer (POE-POB) exhibited favorable selectivity toward VLDLR and IDL fractions. POE-POB removed apolipoprotein (apo) E and apo C-III from IDL particles in the presence of cholesterol esterase (CHER), and the particle size distribution of IDLs became smaller after the reaction. These results revealed that IDL particles are specifically modified in the presence of CHER and POE-POB, making their component cholesterol available for enzymatic assay. Addition of phospholipase D improved the reactivity toward chylomicron remnants (CMRs). We found a high correlation [y = 1.018x- 0.01 mmol/L, r = 0.962 (n = 160)] between the proposed assay and the immunoseparation assay in serum from healthy individuals.Conclusion: The homogeneous assay described in this report can measure TRL remnant cholesterol, including CMRs, VLDLRs, and, IDLs, with high sensitivity and specificity. (C) 2007 American Association for Clinical Chemistry.