Shi Ce, Vogg Sebastian, Lin Dong-Qiang, Sponchioni Mattia, Morbidelli Massimo
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
YMC ChromaCon, Zürich, Switzerland.
Biotechnol Bioeng. 2021 Sep;118(9):3420-3434. doi: 10.1002/bit.27763. Epub 2021 Apr 13.
The increasing demand for efficient and robust processes in the purification of monoclonal antibodies (mAbs) has recently brought frontal chromatography to the forefront. Applied during the polishing step, it enables the removal of high molecular weight aggregates from the target product, achieving high purities. Typically, this process is operated in batch using a single column, which makes it intrinsically subjected to a purity-yield tradeoff. This means that high purities can only be achieved at the cost of lowering the product yield and vice versa. Recently, a two-column continuous implementation of frontal chromatography, referred to as Flow2, was developed. Despite being able of alleviating the purity-yield tradeoff typical of batch operations, the increase in the number of process parameters complicates its optimal design, with the risk of not exploiting its full potential. In this study, we developed an ad hoc design procedure (DP) suitable for the optimization of both batch frontal chromatography and Flow2 in terms of purity, yield, and productivity. This procedure provided similar results as a multiobjective optimization based on genetic algorithm but with lower computational effort. Then, batch and Flow2 operated at their optimal conditions were compared. Besides showing a more favorable Pareto front of yield and productivity at a specified purity, the Flow2 process demonstrated improved robustness compared to the batch process with respect to modifications in the loading linear velocity, washing buffer ionic strength and loading time, thus providing an appealing operation for integrated processes.
近期，对单克隆抗体（mAb）纯化中高效且稳健工艺的需求不断增加，使得前沿色谱法成为了焦点。在精制步骤中应用时，它能够从目标产物中去除高分子量聚集体，从而实现高纯度。通常，此过程采用单柱进行间歇操作，这使其本质上存在纯度 - 收率的权衡。这意味着只能以降低产物收率为代价来实现高纯度，反之亦然。最近，开发了一种两柱连续式前沿色谱法，称为Flow2。尽管它能够缓解间歇操作中典型的纯度 - 收率权衡问题，但工艺参数数量的增加使其优化设计变得复杂，存在无法充分发挥其潜力的风险。在本研究中，我们开发了一种专门的设计程序（DP），适用于在纯度、收率和生产率方面对间歇前沿色谱法和Flow2进行优化。该程序提供的结果与基于遗传算法的多目标优化相似，但计算量更低。然后，对在最佳条件下运行的间歇式和Flow2进行了比较。除了在特定纯度下呈现出更有利的收率和生产率帕累托前沿外，Flow2工艺在进样线性速度、洗涤缓冲液离子强度和进样时间的变化方面，相对于间歇工艺表现出更高的稳健性，从而为集成工艺提供了一种有吸引力的操作方式。
