离子交换法纯化重组类人胶原蛋白II的研究  

PURIFICATION OF RECOMBINANT HUMAN COLLAGEN-LIKE BIOPROTEIN II BY ION EXCHANGE CHROMATOGRAPHY

作　　者：薛文娇[1] 范代娣[1] 尚龙安[1] 侯文洁[1] 鱼奇鳞[2] 张卫刚 

机构地区：[1]西北大学化工学院生物工程系,西安710069 [2]长安大学环境科学与工程学院,西安710061 [3]西安交大保赛生物技术股份有限公司,西安710048

出　　处：《离子交换与吸附》2006年第4期289-297,共9页Ion Exchange and Adsorption

基　　金：国家科技攻关重大专项(2003DA901A32);国家自然科学基金(No.20476085)

摘　　要：用阳离子交换树脂CM-52分离纯化类人胶原蛋白II(HumanCollagen-likeBioproteinII,HCBII)。通过实验,分别确定了静态吸附和柱层析吸附的操作条件,即静态吸附:在pH4.0、NaCl0.15mol/L、进料浓度7g/L,处理量17.26ml/g树脂的条件下进行吸附,吸附时间为60min;柱层析:在pH4.0、NaCl0.15mol/L、进料浓度5g/L、流速5ml/min的条件下进行吸附。然后上柱,在pH4.0、NaCl0.30mol/L下进行脱附。实验表明,采用静态吸附的方式吸附HCBII,然后上柱洗脱,HCBII的吸附量可达到48.6mg/g树脂,回收率83.8%,操作时间短,分辨率高,最终纯化的HCBII达电泳纯,分子量为97kD。A purification method of recombinant human collagen-like Bioprotein Ⅱ （HCB Ⅱ） by the CM52 cation exchange chromatography were reported. The batch operation and normal chromatography column method were used in experiments, i.e., Batch operation of chromatography： HCB Ⅱ was absorbed under pH 4.0, NaCl （ionic strength） 0.15mol/L, 17.26ml （protein volume）/g （resin） and concentration of crude protein liquid 7g/L; Column chromatography： HCB Ⅱ was absorbed under pH 4.0, NaCl （ionic strength） 0.15mol/L, flow rate 5ml/min and concentration of crude protein liquid 5g/L, the elution of HCB Ⅱ was pH 4.0, the concentration of NaCl （ionic strength） 0.30mol/L. As a result, the CM52 cation exchange chromatography was of better capacity of adsorption for HCB Ⅱ and the elution was simple; the total purity process time would be shorten and resolution was high, The recovery of aim protein HCB Ⅱ with a molecular weight around 97kD was above 83.8% and purity of HCB Ⅱ could reach electrophoresis grade purity.

关 键 词：HCBⅡ 离子交换 静态吸附 柱层析吸附 回收率 

分 类 号：TQ028.3[化学工程] TQ936.22