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.
出处
《生物工程学报》
CAS
CSCD
北大核心
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 )~~
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
expended bed adsorption, purification process, contin uous perfusion culture, monoclonal antibody
