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首页> 外文会议>Conference on single-use technologies II: bridging polymer science to biotechnology applications >FLOW-THROUGH CHROMATOGRAPHY AS A CONTINUOUS AND INTEGRATED PURIFICATION METHOD
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FLOW-THROUGH CHROMATOGRAPHY AS A CONTINUOUS AND INTEGRATED PURIFICATION METHOD

机译：连续色谱纯化的流式色谱法

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摘要

Continuous manufacturing is expected to improve the efficiency and economics of protein and other bio-product production processes. However, it is not easy to design and operate the continuous process especially for downstream processing as many unit operations (chromatography and membrane filtration) are involved. An operation method known as flow-through chromatography (FTC) is considered to be an efficient purification method as the flow is continuous. In FTC, a target bio-product is eluted from the chromatography column without adsorption whereas contaminants are strongly bound. Usually two different modes of chromatography are needed in order to remove various kinds of contaminants. Two FTC columns have to be connected in order to build the integrated continuous process. This is not an easy task since the mobile phase properties (pH, salt, buffer ions) are different for the two columns. In this study, we investigated how FTC processes can be designed based on linear gradient elution (LGE) data by using our LGE model. As a first model separation system removal of BSA dimer from BSA monomer was chosen. The distribution coefficients of BSA monomer and dimer and the mass transfer data were obtained from LGE experimental data based on the model. Experimental breakthrough of BSA dimer was well predicted by the model simulation. The model simulation also showed that FTC is very sensitive to a small change in salt concentration and/or pH of the mobile phase as well as the mobile phase velocity (see Fig.1). The productivity calculation method was also developed. The second model system was to use two FTC columns for removing multiple contaminants. In this system the efficiency of FTC processes was examined in terms of impurity removal efficiency from the cell culture broth containing monoclonal antibody. It was found that two FTC (anion exchange chromatography and cation exchange chromatography) can remove impurities efficiently when the mobile phase pH and conductivity were properly chosen. It was also shown that the two columns can be connected as a pseudo continuous FTC operation.
机译：连续生产有望提高蛋白质和其他生物产品生产过程的效率和经济性。但是，设计和操作连续过程并不容易，特别是对于下游过程而言，因为涉及许多单元操作（色谱和膜过滤）。由于流动是连续的，因此称为流通色谱（FTC）的操作方法被认为是一种有效的纯化方法。在FTC中，目标生物产物从色谱柱上洗脱下来而没有吸附，而污染物则被牢固地结合在一起。通常，需要两种不同的色谱模式以去除各种污染物。必须连接两个FTC柱才能构建集成的连续过程。这不是一件容易的事，因为两根色谱柱的流动相特性（pH，盐，缓冲离子）不同。在这项研究中，我们研究了如何使用LGE模型基于线性梯度洗脱（LGE）数据设计FTC过程。作为第一个模型分离系统，选择了从BSA单体中去除BSA二聚体的方法。基于该模型，从LGE实验数据获得了BSA单体和二聚体的分布系数以及传质数据。通过模型仿真可以很好地预测BSA二聚体的实验突破。模型仿真还表明，FTC对流动相盐浓度和/或pH的微小变化以及流动相速度非常敏感（见图1）。还开发了生产率计算方法。第二个模型系统是使用两个FTC色谱柱去除多种污染物。在该系统中，根据从含有单克隆抗体的细胞培养液中去除杂质的效率来检查FTC过程的效率。结果发现，2 FTC（阴离子交换层析和阳离子交换层析）能有效地除去杂质时，流动相的pH和电导率分别适当选择。还显示了两列可以作为伪连续FTC操作连接。

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