一种新颖的除去重组人血清白蛋白杂质的方法  

A Novel Method of Removing Impurities from Recombinant Human Serum Albumin

作　　者：李世杰[1,2] 袁彩[2,3] 郑科[3] 陈锦灿[2] 雪光浦 陈卓[2] 黄明东[2,3] LI Shi-jie YUAN Cai ZHENG Ke CHEN Jin-can XUE Guang-pu CHEN Zhuo HUANG Ming-dong(College of Life Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China State Key Laboratory of Structure Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou 350002, China College of Chemistry, Fuzhou University, Fuzhou 350116, China College of Life Science, Fujian Normal University, Fuzhou 350117, China)

机构地区：[1]福建农林大学生命科学学院,福建福州350002 [2]中国科学院福建物质结构研究所结构化学国家重点实验室,福建福州350002 [3]福州大学化学学院,福建福州350116 [4]福建师范大学生命科学学院,福建福州350117

出　　处：《福建师范大学学报（自然科学版）》2017年第4期65-72,共8页Journal of Fujian Normal University：Natural Science Edition

基　　金：国家自然科学基金资助项目 ( 31170707;31370737;31400637;31570745;31670739)

摘　　要：开发了一种简便高效的去除杂质的方法 (CBIR法).在含6 mol·L-1尿素、体积分数为30%酒精的不含还原剂的体系中,重组人血清白蛋白的结构被部分变性而其二硫键不被破坏,从而使吸附及包裹在其内部的色素等杂质暴露,而与重组人血清白蛋白分离.由于多对二硫键的存在,在去除变性剂情况下,人血清白蛋白恢复原来的构象.通过一系列实验对复性后的人血清白蛋白纯度和活性进行检测,确定这种CBIR法在提高人血清白蛋白纯度的同时,并不影响其活性.此外,也同样适用于重组人血清白蛋白融合蛋白的纯化,具有广泛的适用性.A simple and effective method was developed to remove recombinant human serum albumin（ r HSA） impurities. In the presence of 6 mol·L-1urea and 30% alcohol volume fraction and disulfide bonds are not broken,the structure of r HSA is partial unfolded and disulfide bonds are not broken. The impurities adsorbed or entrapped in r HSA are exposed outside,and can be washed out from r HSA. Upon the removal of protein denaturant,the conformation of r HSA restores quickly owing to the presence of multiple disulfide bonds. It demonstrate that this method improve significantly the purity of r HSA without affecting bioactivity of r HSA. In addition,the method is also used successfully for a number of albumin fusion proteins,demonstrating the wide applicability of this novel method.

关 键 词：重组人血清白蛋白 融合蛋白 变复性 尿素 生物活性 

分 类 号：Q816[生物学—生物工程]