Measurement of prebeta-1 HDL in human plasma by an ultrafiltration-isotope dilution technique.
Measurement of prebeta-1 HDL in human plasma by an ultrafiltration-isotope dilution technique.
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DOI：
10.1006/abio.1997.2258
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发表时间：
1997-09
影响因子：
2.9
通讯作者：
P. O’Connor;J. Naya-Vigne;Philippe Duchateau;B. Ishida;M. Mazur;S. Schoenhaus;B. Zysow;M. Malloy-M.
中科院分区：
文献类型：
--
作者：
P. O’Connor;J. Naya-Vigne;Philippe Duchateau;B. Ishida;M. Mazur;S. Schoenhaus;B. Zysow;M. Malloy-M.
Prebeta-1 HDL是一种67 kDa的血浆高密度脂蛋白（HDL），含有两个载脂蛋白A-I拷贝。它在胆固醇回收的代谢循环中起作用，并可能在血浆中的脂解过程中形成。我们已经发现，离心超滤使用的膜与100 kDa的渗透性限制之间的67 kDa的种类和较大的HDL颗粒种类的区分。因此，超滤液对血浆中的前β-1 HDL池进行采样。我们已经开发了一种技术，使用纯化的prebeta-1 HDL的分散，用氚共价标记，在血浆样品中，标记的prebeta-1 HDL池。随后测定超滤液中前β-1 HDL的比活度，提供了通过同位素稀释原理计算血浆中前β-1 HDL含量的方法。我们采用酶联免疫吸附测定技术的修改载脂蛋白A-I，允许该蛋白在prebeta-1 HDL和在其他HDL颗粒物种的测定总载脂蛋白A-I的分数是存在于prebeta-1 HDL颗粒物种的相等检测。86名血脂正常成人血浆样本中前β 1 HDL相关载脂蛋白A-I的平均水平为74 +/- 43 μ g/ml（+/-SD），平均占血浆总载脂蛋白A-I的6.6%。
Prebeta-1 HDL is a 67-kDa species of plasma high-density lipoproteins (HDL) that contains two copies of apolipoprotein A-I. It functions in a metabolic cycle of cholesterol retrieval and may be formed during lipolysis in plasma. We have found that centrifugal ultrafiltration using a membrane with a permeability limit of 100 kDa discriminates categorically between the 67-kDa species and larger HDL particle species. Thus, the ultrafiltrate samples the pool of prebeta-1 HDL in plasma. We have developed a technique using the dispersal of purified prebeta-1 HDL, labeled covalently with tritium, in plasma samples, to label the prebeta-1 HDL pool. Subsequent determination of the specific activity of prebeta-1 HDL in the ultrafiltrate provides a means of calculating the content of prebeta-1 HDL in plasma by the isotope dilution principle. We employ a modification of an enzyme-linked immunosorbent assay technique for apolipoprotein A-I that allows the equal detection of that protein in prebeta-1 HDL and in other HDL particle species for determination of the fraction of total apolipoprotein A-I that is present in the prebeta-1 HDL particle species. The mean level of prebeta-1 HDL-associated apolipoprotein A-I in plasma samples from 86 normolipidemic adults was 74 +/- 43 microg/ml (+/-SD), representing an average of 6.6% of the total apolipoprotein A-I in plasma.