摘要
目的：分析单克隆抗体纯化过程中，去除内毒素的不同方法的应用效果，并探讨它们应用于中试规模的可行性。方法与结果：比较了多聚赖氨酸型的内毒素去除填料、20nm膜过滤、将单抗附着在蛋白A柱上后使用精氨酸和组氨酸溶液冲洗等3种方法的内毒素去除效果，发现3种方法都可以将内毒素水平大幅降低，可分别将内毒素去除70％、88％和97％。因为单抗分子等电点较高，所获得的最低内毒素含量为0．2～0-3EU／mg。结论：3种方法均具有一定的工艺放大潜力，进一步提高内毒素去除效果将需要综合使用不同机理的去除技术。
Objective： To study the effectiveness of three different methods to reduce endotoxin levels during pu- rified monoclonal antibody（mAb） preparations. Methods ＆ Results： Commercial ploy-lysine conjugated endotoxin removal resin, 20 nm membrane fihration, as well as arginine and histidine solution washing of mAb captured in protein A column were used to reduce endotoxin levels. All three methods were capable of significantly reducing endotoxin content by 70%, 88%, and 97%, respectively. However, since the mAb may be positively charged due to high pI value, minimum endotoxin level achieved was at 0.2-0.3 EU/mg range. Conclusion： All three methods have potential to be scalable in the mAb purification process. However, further improvement of endotoxin removal efficiency may rely upon combination of methods with different mechanism.
出处
《生物技术通讯》
CAS
2013年第1期94-97,142,共5页
Letters in Biotechnology
关键词
内毒素去除
单克隆抗体纯化
组氨酸
精氨酸
eudotoxin removal
monoclonal antibody purification
arginine
histidine
