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Biotechnol J. 2019 Apr;14(4). doi: 10.1002/biot.201800243. Epub 2018 Nov 27.
An emphasis on continuous monoclonal antibody (mAb) bioprocessing in the pharmaceutical industry necessitates effective approaches for downstream process development (PD). With a PD strategy, the process parameters are optimized to directly develop streamlined three-step continuous chromatography processes. A design of experiment (DoE) approach with single column (batch mode) is used to simulate a multi-column (continuous mode) purification method and characterize each chromatography step: Protein A capture, anion exchange, and mixed mode cation exchange. A novel and targeted approach to quickly characterize a DoE design space was employed and empirical modeling was used to define the capacity for multi-column chromatography to accurately transfer the batch process to continuous mode of purification. This PD approach effectively mimics the continuous mode of operation and provides the flexibility to develop multiple continuous processes with target mAb recovery and purity. By implementing this PD strategy and the process parameters defined in batch mode, two robust and predictable continuous bioprocesses were developed within 7 weeks of investigation, which resulted in a total product yield of recovery at or above 74%, host cell protein (HCP) content below 5 ppm, and aggregate content below 1%.
在制药行业中，对连续单克隆抗体（mAb）生物工艺的强调需要有效的下游工艺开发（PD）方法。通过 PD 策略，可以优化工艺参数，直接开发简化的三步连续色谱工艺。采用单柱（批处理模式）的实验设计（DoE）方法来模拟多柱（连续模式）纯化方法，并对每个色谱步骤进行表征：Protein A 捕获、阴离子交换和混合模式阳离子交换。采用了一种新颖的、有针对性的方法来快速表征 DoE 设计空间，并使用经验模型来定义多柱色谱的容量，以将批处理过程准确地转移到连续纯化模式。这种 PD 方法有效地模拟了连续操作模式，并提供了开发具有目标 mAb 回收率和纯度的多个连续工艺的灵活性。通过实施这种 PD 策略和批处理模式中定义的工艺参数，在 7 周的研究时间内开发了两个稳健且可预测的连续生物工艺，总产品回收率达到或高于 74%，宿主细胞蛋白（HCP）含量低于 5ppm，聚集体含量低于 1%。
