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通过使用单克隆抗体的酶联免疫吸附测定法定量脂蛋白(a)。

Lipoprotein(a) quantified by an enzyme-linked immunosorbent assay with monoclonal antibodies.

作者信息

Labeur C, Michiels G, Bury J, Usher D C, Rosseneu M

机构信息

Department of Clinical Chemistry, Brugge, Belgium.

出版信息

Clin Chem. 1989 Jul;35(7):1380-4.

PMID:2527093
Abstract

This new, sensitive, specific "sandwich"-type enzyme-linked immunosorbent assay (ELISA) for quantifying lipoprotein(a) [Lp(a)] in human serum and in ultracentrifugal lipoprotein fractions is based on use of a monoclonal antibody raised against apolipoprotein(a) as coating protein and a polyclonal antibody, raised against either apo B or against Lp(a) and conjugated with peroxidase, for detection of bound Lp(a). Mean intra- and interassay CVs for assay of 16 samples were 3.0% and 5.6%, respectively. Sample pretreatment with urea did not enhance Lp(a) immunoreactivity, and treatment with nonionic detergents decreased binding to the monoclonal antibody. Results correlated well (r = 0.99, n = 38) with those by radial immunodiffusion (RID). The ELISA assay, however, detects amounts corresponding to Lp(a) contents of 10 to 1000 mg/L in plasma samples diluted 1000-fold, compared with 100-500 mg/L for RID. For 92 normolipidemic subjects, the mean Lp(a) concentration was 120 (SD 130) mg/L. In patients undergoing coronary angiography, Lp(a) concentrations increased with the severity of the disease but were not correlated with either HDL cholesterol, triglycerides, apo A-I, or apo B, and only weakly with plasma cholesterol and apo A-II. These two correlations were even weaker in normal subjects, and only the correlation with total cholesterol was valid. Lp(a), measured at birth and at seven days and six months, steadily increased with age. This assay is well suited for measuring Lp(a) in plasma and in lipoprotein fractions and also for screening programs evaluating this significant genetic risk factor for the development of atherosclerosis.

摘要

这种新型、灵敏、特异的“夹心”型酶联免疫吸附测定法(ELISA)用于定量检测人血清及超速离心脂蛋白组分中的脂蛋白(a)[Lp(a)],其原理是使用针对载脂蛋白(a)产生的单克隆抗体作为包被蛋白,以及针对载脂蛋白B或Lp(a)产生的、与过氧化物酶偶联的多克隆抗体来检测结合的Lp(a)。对16个样本进行测定的平均批内和批间变异系数分别为3.0%和5.6%。用尿素对样本进行预处理不会增强Lp(a)的免疫反应性,用非离子去污剂处理会降低与单克隆抗体的结合。结果与放射免疫扩散法(RID)的结果相关性良好(r = 0.99,n = 38)。然而,ELISA测定法在将血浆样本稀释1000倍后,能检测出相当于10至1000 mg/L Lp(a)含量的量,而RID法为100 - 500 mg/L。对于92名血脂正常的受试者,平均Lp(a)浓度为120(标准差130)mg/L。在接受冠状动脉造影的患者中,Lp(a)浓度随疾病严重程度增加,但与高密度脂蛋白胆固醇、甘油三酯、载脂蛋白A - I或载脂蛋白B均无相关性,与血浆胆固醇和载脂蛋白A - II的相关性也较弱。在正常受试者中,这两种相关性更弱,只有与总胆固醇的相关性是有效的。在出生时、7天和6个月时测量的Lp(a)随年龄稳步增加。该测定法非常适合测量血浆和脂蛋白组分中的Lp(a),也适用于评估这种动脉粥样硬化发展的重要遗传风险因素的筛查项目。

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