【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。更多还原

【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. ApoCⅠ,CⅡ and CⅢ were purified from apoCs fraction by DEAE-Sephacel ion exchange chromatography. Results Purified apoE,apoCⅠ,apoCⅡ and apoCⅢ 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.更多还原