三步法制备级纯化抗CD20(Fab′)_2  

Three-step Purification of Preparative-scale AntiCD20(Fab′)_2

作　　者：王金宏[1] 杨铭[1] 范冬梅[1] 许元生[1] 熊冬生[1] 杨纯正[1] 

机构地区：[1]中国医学科学院北京协和医学院血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《中国医学科学院学报》2008年第5期622-625,共4页Acta Academiae Medicinae Sinicae

基　　金：天津市科技发展计划项目(05YFGZGX02800)~~

摘　　要：目的在小规模蛋白纯化系统AKTA prime上建立制备级纯化抗CD20(Fab′)2的方法。方法将通过高渗溶液提取的周质腔蛋白抗CD20(Fab′)2依次经过离子柱、疏水柱和亲和柱纯化,采用蛋白电泳和高效液相色谱法分析检测其分离效果及纯度,同时测定其与Raji细胞的结合活性。结果在该纯化条件下一次可获得8mg纯度为96.678%的抗CD20(Fab′)2,其与CD20+Raji细胞的结合活性与采用亲和柱联合分子筛柱得到的抗体活性基本一致。结论三步法制备级纯化抗CD20(Fab′)2操作简单,能获得制备级高纯度抗CD20(Fab′)2。Objective To establish a three-step purification method of preparative-scale antiCD20 （Fab＇） ： using AKTA prime. Methods AntiCD20 （Fab＇）2 was extracted by hyperosmotic solution and then purified by CM sepharose FF, phenyl sepharose FF, and protein G sepharose FF. Results Around 8 mg anti- CD20 （ Fab＇ ） 2, whose purification was 96. 678% , was purified. The antigen-binding activity of antiCD20 （Fab＇）2 was similar to that of antiCD20 （Fab＇）2 purified by protein G sepharose FF and S-100. Conclusion The three-step purification method can obtain high-purity preparative- scale antiCD20 （Fab＇）2 in a simple way.

关 键 词：抗CD20(Fab′)2 三步法纯化 高效液相色谱分析 

分 类 号：R963[医药卫生—微生物与生化药学]