单克隆抗体纯化过程中内毒素去除方法分析

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。

Investigation of Methods for Endotoxin Removal in Monoclonal Antibody Purification Process

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.