摘要
用密度梯度超速离心法，从混合人血清中分离极低密度脂蛋白，经脱脂后再进行聚焦层析分离其中的载脂蛋白。获得的纯品载脂蛋白ＣⅢ１和ＣⅢ２经等电聚焦电泳均呈现一条带。生物活性的细胞学实验鉴定发现，纯化载脂蛋白ＣⅢ能抑制培养ＨｅｐＧ２细胞对标记极低密度脂蛋白的提取。聚焦层析方法简便，一次上样量大，速度快，分离效果好，还可同时获得载脂蛋白ＣⅠ，ＣⅡ和Ｅ。
This study aimed to isolate isoforms of apolipoprotein (apo) CⅢ from human serum. 24 hour fasting serum from normal and hyperlipidemic subjects was pooled and subjected to ultracentrifugation at plasma density for 20 hours. Very low density lipoprotein (VLDL) was collected at density of d<1.006g/ml, and it was delipidated by ethanol and ether. The delipidated apo VLDL was dissolved in a solution containing 7.2mol/L urea and 20mmol/L dithiothreitol. The insoluble apo B was removed by centrifugation. The soluble apo VLDL was applied to PBE94 column, and eluted with elution buffer containing polybuffer 74 and 8mol/L urea (1∶8, pH4.0). After pooled, the eluted peaks of apolipoproteins were applied to column chromatography of hydroxylapatite to remove the polybuffer. The purified isoforms of apoC Ⅲ and the purified apo CⅠ, CⅡand E, were characterized by isoelectrofocusing and west blot. The results showed that the purified apoCⅢ 1, CⅢ 2, and CⅡ were pure.
出处
《华西医科大学学报》
CSCD
1999年第1期111-113,共3页
Journal of West China University of Medical Sciences
