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陶瓷羟基磷灰石广泛用于各种生物分子的分离和纯化,但在层析应用上,纯化洗脱过程中填料表面氢原子的释放容易导致其性能和寿命降低。研究了一种可以延长CHT填料使用寿命的方法,通过在洗脱步骤前加入一种称为表面中和体系(SNS)的溶液,结合释放的氢原子,使得整个体系处于一个接近中性pH的状态。先筛选出有效调整pH的SNS条件,通过钙离子流失的监测证实该条件的有效性。再通过在WLB304单克隆抗体纯化工艺中加入SNS之后,对羟基磷灰石填料载量、pH和压力的变化、杂质的去除能力进行考察。结果表明,加入SNS可以有效减少钙离子流失,同时并不会影响CHT层析工艺的效果。
Ceramic hydroxyapatite is widely used in the separation and purification of various biomolecules. However, in the chromatographic applications, the release of hydrogen atoms on the surface of the packing during purification and elution easily leads to the decrease of its performance and life span. A method of extending the service life of a CHT filler was studied by adding a solution called Surface Neutralization System (SNS) prior to the elution step in combination with the release of hydrogen atoms to bring the overall system to a near neutral pH status. SNS conditions for effectively adjusting pH were screened first, and the validity of this condition was confirmed by the monitoring of calcium loss. By adding SNS to WLB304 monoclonal antibody purification process, the loading, pH and pressure of hydroxyapatite, the removal of impurities were investigated. The results show that the addition of SNS can effectively reduce the loss of calcium, while not affecting the CHT chromatography process.