Institute for Chemical and Bioengineering, ETH Zurich, Zurich, Switzerland.
Biotechnol Bioeng. 2010 Dec 15;107(6):974-84. doi: 10.1002/bit.22887.
A two-step chromatography process for monoclonal antibody (mAb) purification from clarified cell culture supernatant (cCCS) was developed using cation exchange Multicolumn Countercurrent Solvent Gradient Purification (MCSGP) as a capture step. After an initial characterization of the cell culture supernatant the capture step was designed from a batch gradient elution chromatogram. A variety of chromatographic materials was screened for polishing of the MCSGP-captured material in batch mode. Using multi-modal anion exchange in bind-elute mode, mAb was produced consistently within the purity specification. The benchmark was a state-of-the-art 3-step chromatographic process based on protein A, anion and cation exchange stationary phases. The performance of the developed 2-step process was compared to this process in terms of purity, yield, productivity and buffer consumption. Finally, the potential of the MCSGP process was investigated by comparing its performance to that of a classical batch process that used the same stationary phase.
两步层析法从澄清的细胞培养上清液（cCCS）中纯化单克隆抗体（mAb），采用阳离子交换多柱逆流溶剂梯度纯化（MCSGP）作为捕获步骤。在对细胞培养上清液进行初步表征后，从批梯度洗脱色谱图中设计了捕获步骤。筛选了多种色谱材料，以在批处理模式下对 MCSGP 捕获的材料进行抛光。使用多模式阴离子交换在结合洗脱模式下，mAb 的纯度始终符合规范。基准是基于蛋白 A、阴离子和阳离子交换固定相的 3 步色谱工艺。从纯度、收率、生产率和缓冲液消耗等方面比较了开发的 2 步工艺与该工艺的性能。最后，通过比较相同固定相的经典批处理过程的性能，研究了 MCSGP 工艺的潜力。
