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。
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.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2007年第2期328-330,333,共4页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(批准号398702298)资助
