Beck J T, Williamson B, Tipton B
AMGEN, Thousand Oaks, CA 91320-1789, USA.
Bioseparation. 1999;8(1-5):201-7.
In the course of developing a cost-effective, scaleable process for the purification of a recombinant protein from Chinese hamster ovary (CHO) suspension cell culture, we investigated direct capture of this molecule using expanded bed adsorption (EBA). EBA combines clarification, purification, and concentration of the product into a single step. The unclarified bioreactor material was directly applied to a STREAMLINE 25 column containing an affinity STREAMLINE adsorbent. This work focused on simplifying the EBA operations and minimizing the overall processing time by running the EBA column unidirectionally, eluting in the expanded bed mode, and coupling the EBA column directly with ion exchange or hydrophobic interaction chromatography. Unidirectional EBA was clearly a simpler unit operation and did not require the use of specialized equipment. The increase in the elution pool volume was insignificant, especially when the EBA column was eluted directly onto the downstream column. Scale-down was simple and could be automated. Coupling of unidirectional EBA with a downstream purification step reduced processing time, equipment requirements and cost.
在开发一种从中国仓鼠卵巢（CHO）悬浮细胞培养物中纯化重组蛋白的经济高效、可扩展的工艺过程中，我们研究了使用扩张床吸附（EBA）直接捕获该分子的方法。EBA将产物的澄清、纯化和浓缩整合为一个步骤。未澄清的生物反应器物料直接应用于装有亲和STREAMLINE吸附剂的STREAMLINE 25柱。这项工作的重点是通过单向运行EBA柱、在扩张床模式下洗脱以及将EBA柱直接与离子交换或疏水相互作用色谱联用，简化EBA操作并最大限度地减少整体处理时间。单向EBA显然是一种更简单的单元操作，不需要使用专门设备。洗脱池体积的增加微不足道，尤其是当EBA柱直接洗脱到下游柱上时。缩小规模很简单且可以自动化。单向EBA与下游纯化步骤联用可减少处理时间、设备需求和成本。
