Roberts D C, Miller N E, Price S G, Crook D, Cortese C, La Ville A, Masana L, Lewis B
Biochem J. 1985 Feb 15;226(1):319-22. doi: 10.1042/bj2260319.
A simple method has been developed for labelling human plasma lipoproteins to high specific radioactivity with radioactive cholesteryl esters in vitro. After isolation by preparative ultracentrifugation, the selected lipoprotein was incubated for 30 min at 4 degrees C in human serum (d greater than 1.215) that had been prelabelled with [4-14C]cholesteryl oleate or [1,2-3H]cholesteryl linoleate, and was then re-isolated by ultracentrifugation. All major lipoprotein classes were labelled by the procedure. Specific radioactivities of up to 18 d.p.m. . pmol-1 (46 d.p.m. . ng-1) were achieved. When radiolabelled high-density lipoprotein was infused intravenously, the radioactive cholesteryl ester behaved in vivo indistinguishably from endogenous cholesteryl esters produced by the lecithin (phosphatidylcholine): cholesterol acyltransferase reaction.
已开发出一种简单方法,可在体外将人血浆脂蛋白用放射性胆固醇酯标记至高比放射性。通过制备性超速离心分离后,将选定的脂蛋白在4℃下于已用[4-¹⁴C]胆固醇油酸酯或[1,2-³H]胆固醇亚油酸酯预标记的人血清(d大于1.215)中孵育30分钟,然后通过超速离心重新分离。所有主要脂蛋白类别均通过该程序进行标记。比放射性可达18 d.p.m.·pmol⁻¹(46 d.p.m.·ng⁻¹)。当静脉内注入放射性标记的高密度脂蛋白时,放射性胆固醇酯在体内的行为与卵磷脂(磷脂酰胆碱):胆固醇酰基转移酶反应产生的内源性胆固醇酯无明显区别。