Department of Biotechnology, Delft University of Technology, van der Maasweg 9, Delft, 2629 HZ, the Netherlands.
Downstream Processing, Byondis B.V., Microweg 22, 6503 GB, Nijmegen, the Netherlands; Bioprocessing Engineering, Wageningen University, Droevendaalse steeg 1, 6708 PB, Wageningen, the Netherlands.
J Chromatogr A. 2024 Feb 22;1717:464672. doi: 10.1016/j.chroma.2024.464672. Epub 2024 Jan 23.
The monoclonal antibody (mAb) industry is becoming increasingly digitalized. Digital twins are becoming increasingly important to test or validate processes before manufacturing. High-Throughput Process Development (HTPD) has been progressively used as a tool for process development and innovation. The combination of High-Throughput Screening with fast computational methods allows to study processes in-silico in a fast and efficient manner. This paper presents a hybrid approach for HTPD where equal importance is given to experimental, computational and decision-making stages. Equilibrium adsorption isotherms of 13 protein A and 16 Cation-Exchange resins were determined with pure mAb. The influence of other components in the clarified cell culture supernatant (harvest) has been under-investigated. This work contributes with a methodology for the study of equilibrium adsorption of mAb in harvest to different protein A resins and compares the adsorption behavior with the pure sample experiments. Column chromatography was modelled using a Lumped Kinetic Model, with an overall mass transfer coefficient parameter (k). The screening results showed that the harvest solution had virtually no influence on the adsorption behavior of mAb to the different protein A resins tested. k was found to have a linear correlation with the sample feed concentration, which is in line with mass transfer theory. The hybrid approach for HTPD presented highlights the roles of the computational, experimental, and decision-making stages in process development, and how it can be implemented to develop a chromatographic process. The proposed white-box digital twin helps to accelerate chromatographic process development.
单克隆抗体 (mAb) 行业正变得越来越数字化。在制造之前，数字孪生对于测试或验证工艺变得越来越重要。高通量工艺开发 (HTPD) 已逐渐成为工艺开发和创新的工具。高通量筛选与快速计算方法的结合允许以快速有效的方式对过程进行计算机模拟研究。本文提出了一种 HTPD 的混合方法，其中对实验、计算和决策阶段给予同等重视。使用纯 mAb 确定了 13 种蛋白 A 和 16 种阳离子交换树脂的平衡吸附等温线。澄清的细胞培养上清液 (收获物) 中其他成分的影响尚未得到充分研究。这项工作为研究 mAb 在收获物中对不同蛋白 A 树脂的平衡吸附提供了一种方法，并比较了与纯样品实验的吸附行为。使用总体传质系数参数 (k) 对柱层析进行了集总动力学模型模拟。筛选结果表明，收获溶液对不同蛋白 A 树脂吸附 mAb 的行为几乎没有影响。k 与样品进料浓度呈线性相关，这与质量传递理论一致。所提出的 HTPD 混合方法强调了计算、实验和决策阶段在工艺开发中的作用，以及如何实施该方法来开发色谱工艺。所提出的白盒数字孪生有助于加速色谱工艺开发。
