Baginsky M L, Brown W V
J Lipid Res. 1979 May;20(4):548-56.
Lipoprotein lipase (LPL) and hepatic triglyceride lipase (H-TGL) are lipolytic activities found in postheparin plasma. A simple and precise method for the direct determination of LPL in postheparin plasma is described. Pre-incubations of this plasma (45--60 min at 26 degrees C) with sodium dodecyl sulfate (35--50 mM) in 0.2 M Tris-HCl buffer, pH 8.2, results in the inactivation of H-TGL, while leaving LPL fully active. Direct determination of H-TGL is done in a separate aliquot of the same postheparin plasma sample using previously reported assay conditons that do not measure LPL. The sodium dodecyl sulfate-resistant lipolytic activity has the characteristics of LPL as judged by a) its activation by serum and by apolipoprotein C-II; b) its inactivation (over 90%) by 0.75 M NaCl; and c) its inactivation by a specific antiserum. No sodium dodecyl sulfate-resistant activity was found in postheparin plasma from a patient with LPL deficiency (primary type I hyperlipoproteinemia). An excellent correlation of values was obtained (r = 0.99) for 30 samples assayed after sodium dodecyl sulfate treatment and after immuno-inactivation of H-TGL. The intra-assay coefficient of variation was +/- 11% and 4% before and after normalization of values, respectively.
脂蛋白脂肪酶(LPL)和肝甘油三酯脂肪酶(H-TGL)是在肝素后血浆中发现的脂解活性。本文描述了一种直接测定肝素后血浆中LPL的简单而精确的方法。将该血浆在0.2M Tris-HCl缓冲液(pH 8.2)中与十二烷基硫酸钠(35-50 mM)预孵育(26℃下45-60分钟),可使H-TGL失活,而LPL仍保持完全活性。使用先前报道的不检测LPL的测定条件,在同一肝素后血浆样品的另一等分试样中直接测定H-TGL。通过以下几点判断,耐十二烷基硫酸钠的脂解活性具有LPL的特征:a)其被血清和载脂蛋白C-II激活;b)其被0.75M NaCl灭活(超过90%);c)其被特异性抗血清灭活。在患有LPL缺乏症(原发性I型高脂蛋白血症)的患者的肝素后血浆中未发现耐十二烷基硫酸钠的活性。对30个样品在十二烷基硫酸钠处理后和H-TGL免疫灭活后进行测定,得到了极好的相关性(r = 0.99)。在数值标准化前后,测定内变异系数分别为+/- 11%和4%。