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| 中压液相层析法纯化人VLDL主要载脂蛋白 | |
| 引用本文： | 辛艳飞,段云,邓祖跃,张祖辉,刘秉文. 中压液相层析法纯化人VLDL主要载脂蛋白[J]. 四川大学学报(医学版), 2007, 38(2): 328-330,333 |
| 作者姓名： | 辛艳飞 段云 邓祖跃 张祖辉 刘秉文 |
| 作者单位： | 四川大学华西基础医学与法医学院,载脂蛋白研究室,成都,610041 |
| 摘 要： | 目的 建立中压液相层析分离法纯化人VLDL主要载脂蛋白(apo) E、CⅠ、CⅡ和CⅢ的方法.方法 以低温乙醇法制备人血浆白蛋白后的残余组分FⅡ Ⅳ为原料,经一次性密度梯度超离心得到VLDL.脱脂后的VLDL(apoVLDL)经中压Sephacryl S-200柱层析得到apoE和apoCs粗品.apoE经肝素琼脂糖CL-6B亲和层析进一步纯化,apoCs经DEAE-Sephacel离子交换层析进行分离.结果 apoE经肝素琼脂糖CL-6B亲和层析进一步纯化,可得纯度为93.8%的纯品;apoCs经离子交换层析分离得到纯度为92%以上的apoCⅡ、apoCⅢ.所纯化各载脂蛋白apo E、CⅠ、CⅡ及CⅢ经免疫单扩和SDS-PAGE鉴定为免疫纯及电泳纯.结论 成功建立了一种可大量、快速、高分辨分离提纯载脂蛋白apo E、CⅠ、CⅡ及CⅢ的方法. |
| 关 键 词： | 载脂蛋白E 载脂蛋白CⅠ 载脂蛋白CⅡ 载脂蛋白CⅢ 中压液相层析 液相 层析法 化人 VLDL 载脂蛋白 Liquid Chromatography Middle Pressure Apolipoproteins Human 分离提纯 高分辨 快速 电泳 免疫 apoC 层析分离 离子交换层析 结果 行分离 纯化 |
| 收稿时间： | 2006-05-19 |
| 修稿时间： | 2006-11-08 |
Purification of Human VLDL Apolipoproteins by Middle Pressure Liquid Chromatography |
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| XIN Yan-fei,DUAN Yun,DENG Zu-yue,ZHANG Zu-hui,LIU Bing-wen. Purification of Human VLDL Apolipoproteins by Middle Pressure Liquid Chromatography[J]. Journal of Sichuan University. Medical science edition, 2007, 38(2): 328-330,333 | |
| Authors: | XIN Yan-fei DUAN Yun DENG Zu-yue ZHANG Zu-hui LIU Bing-wen |
| Affiliation: | Apolipoprotein Research Unit, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China. |
| Abstract: | OBJECTIVE: To purify human VLDL apolipoproteins by middle-pressure liquid chromatography. METHODS: Human VLDLs were isolated by one step density ultracentrifugation. Delipided human VLDL was separated by Sephacryl S-200 molecular sieve chromatography. ApoE was purified by heparin Sepharose CL-6B affinity chromatography. ApoC I ,C II and C III were purified from apoC. fraction by DEAE-Sephacel ion exchange chromatography: RESULTS: Purified apoE, apoC I, apoC II and apoC III were obtained. SDS-PAGE and immunodiffusion tests indicated the isolated proteins were pure. CONCLUSION: We have established a purification procedure for human VLDL apolipoproteins with highly efficiency and simplicity by MPLC. |
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