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Adsorptive filtration: A case study for early impurity reduction in an Escherichia coli production process

机译：吸附过滤：大肠杆菌生产过程中早期杂质减少的案例研究

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摘要

Primary recovery of intracellular products from Escherichia coli requires cell disruption which leads to a massive release of process-related impurities burdening subsequent downstream process (DSP) unit operations. Especially, DNA and endotoxins challenge purification operations due to their size and concentrations. Consequently, an early reduction in impurities will not only simplify the production process but also increase robustness while alleviating the workload afterward. In the present work, we studied the proof of concept whether a nonwoven anion exchange filter material decreases soluble impurities immediately at the clarification step of E. coli DSP. In a first attempt, endotoxin burden was reduced by 4.6-fold and the DNA concentration by 3.6-fold compared to conventional depth filtration. A design of experiment for the adsorptive filtration approach was carried out to analyze the influence of different critical process parameters (CPPs) on impurity reduction. We showed that depending on the CPPs chosen, a DNA lowering of more than 3 log values, an endotoxin decrease of approximately 7 logs, and a minor HCP clearance of at least 0.3 logs could be achieved. Thus, we further revealed a chromatography column protecting effect when using adsorptive filtration beforehand.
机译：来自大肠杆菌的细胞内产物的初次恢复需要细胞破坏，这导致巨大释放的过程相关的杂质负担随后的下游过程（DSP）单元操作。特别是，DNA和内毒素由于其尺寸和浓度而攻击净化操作。因此，杂质的早期降低不仅将简化生产过程，而且还增加了鲁棒性，同时减轻了后续工作量。在目前的工作中，我们研究了无论是非织造阴离子交换过滤器材料在大肠杆菌DSP的澄清步骤中是否立即降低可溶性杂质的概念证明。在第一次尝试中，与常规深度过滤相比，内毒素负荷降低了4.6倍，DNA浓度将3.6倍下降3.6倍。进行了吸附过滤方法的实验设计，以分析不同关键工艺参数（CPP）对杂质减少的影响。我们表明，根据所选择的CPP，降低超过3个数值的DNA，内毒素减小约7个日志，并且可以实现至少0.3个日志的次要HCP清除。因此，我们进一步揭示了在预先使用吸附过滤时的色谱柱保护效果。

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