Pirrung Silvia M, Parruca da Cruz Diogo, Hanke Alexander T, Berends Carmen, Van Beckhoven Ruud F W C, Eppink Michel H M, Ottens Marcel
Dept. of Biotechnology, Delft University of Technology, Van der Maasweg 9, Delft, 2629 HZ, the Netherlands.
DSM Biotechnology Center, Alexander Fleminglaan 1, AX Delft, 2613, the Netherlands.
Biotechnol Prog. 2018 Jul;34(4):1006-1018. doi: 10.1002/btpr.2642. Epub 2018 Jul 1.
The application of mechanistic models for chromatography requires accurate model parameters. Especially for complex feedstocks such as a clarified cell harvest, this can still be an obstacle limiting the use of mechanistic models. Another commonly encountered obstacle is a limited amount of sample material and time to determine all needed parameters. Therefore, this study aimed at implementing an approach on a robotic liquid handling system that starts directly with a complex feedstock containing a monoclonal antibody. The approach was tested by comparing independent experimental data sets with predictions generated by the mechanistic model using all parameters determined in this study. An excellent agreement between prediction and experimental data was found verifying the approach. Thus, it can be concluded that RoboColumns with a bed volume of 200 μL can well be used to determine isotherm parameters for predictions of larger scale columns. Overall, this approach offers a new way to determine crucial model input parameters for mechanistic modelling of chromatography for complex biological feedstocks. © 2018 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:1006-1018, 2018.
色谱机理模型的应用需要准确的模型参数。特别是对于诸如澄清的细胞收获液等复杂原料，这仍然可能是限制机理模型使用的一个障碍。另一个常见的障碍是用于确定所有所需参数的样品材料和时间有限。因此，本研究旨在在机器人液体处理系统上实施一种方法，该方法直接从含有单克隆抗体的复杂原料开始。通过将独立的实验数据集与使用本研究中确定的所有参数的机理模型生成的预测进行比较，对该方法进行了测试。发现预测结果与实验数据之间具有极好的一致性，从而验证了该方法。因此，可以得出结论，床体积为200 μL的RoboColumns可很好地用于确定等温线参数，以预测更大规模的色谱柱。总体而言，这种方法为确定复杂生物原料色谱机理建模的关键模型输入参数提供了一种新途径。© 2018作者 由Wiley Periodicals, Inc.代表美国化学工程师学会出版的《生物技术进展》 生物技术进展，34:1006 - 1018，2018年。
