CAJ | 학술논문

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、20 nm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0.2~0.3 EU/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.
목적:분석단극륭항체순화과정중,거제내독소적불동방법적응용효과,병탐토타문응용우중시규모적가행성.방법여결과:비교료다취뢰안산형적내독소거제전료、20 nm막과려、장단항부착재단백A주상후사용정안산화조안산용액충세등3충방법적내독소거제효과,발현3충방법도가이장내독소수평대폭강저,가분별장내독소거제70%、88%화97%.인위단항분자등전점교고,소획득적최저내독소함량위0.2~0.3 EU/mg.결론:3충방법균구유일정적공예방대잠력,진일보제고내독소거제효과장수요종합사용불동궤리적거제기술.
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 filtration, 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.