扩张柱床吸附层析回收纯化灌流培养生产的单克隆抗体  

Recovery and Purification of Monoclonal Antibody Producted byContinuous Perfusion Culture Using Expended Bed Adsorption

作　　者：余晓玲[1] 米力[1] 姚西英[1] 陈志南[1] 

机构地区：[1]第四军医大学细胞工程研究中心,国家863计划西安细胞工程基地,陕西西安710032

出　　处：《生物工程学报》2004年第2期227-232,共6页Chinese Journal of Biotechnology

基　　金：国家 8 6 3计划生物工程主题项目基金资助(No .2 0 0 2AA 2 1 70 1 1 )~~

摘　　要：用扩张柱床吸附层析技术 ,一步回收纯化连续灌流培养的单克隆抗体。用StreamlineSP阳离子交换介质在固定床柱XK16 2 0上进行条件摸索 ,扩张床柱Streamline 2 5和 50分别用于小规模条件优化和中试规模放大。培养液中的低浓度单抗经此步处理 ,浓缩 10倍以上 ,纯度提高 5～ 7倍 ,回收率 >90 % ,制备周期比固定柱床层析缩短一半以上。根据培养液中单抗浓度的不同 ,一次处理量为 18～ 50L ,纯化规模由实验室水平 (40 0mg)扩大至中试水平(2g) ,生产成本和工艺复杂性大为降低。应用扩张柱床吸附层析技术 ,建立单克隆抗体回收纯化工艺 ,具有经济、简便。Monoclonal antibody producted by continuous perfusion culture was recovered an d purified by expended bed adsorption chromatography. A packed bed column XK16/2 0 was used for method scouting with Streamline SP adsorbent. Two expended bed c olumns Streamline-25 and -50 were used for method optimization and pilot scale experiment,respectively. The recovery yield of monoclonal antibody was above 90 % in a 5～7 fold enhanced purity and 10 fold increased concentration. According to the different concentration of monoclonal antibody in cell culture broth,abou t18～50L fluid can be treated in a single cycle. MAb purification from lab scale (400mg per cycle) to a small pilot scale (2g per cycle) has been achieved.Compa red with packed bed adsorption, the preparation cycle was half shortened, and th e cost of production and the complexity of process were decreased markedly. It h as been proven that a purification process based on expended bed adsorption tech nique is simple, efficient and economical.

关 键 词：扩张柱床吸附层析技术 纯化工艺 灌流培养 单克隆抗体 培养液 

分 类 号：R39233[医药卫生—免疫学]