On-column virus inactivation by solvent/detergent treatment for a recombinant biological product

下游加工 色谱法 化学 重组DNA 产量(工程) 病毒 过程(计算) 生物化学 病毒学 生物 材料科学 计算机科学 冶金 基因 操作系统
作者
Daniel Polasek,Andreas Flicker,Christian Fiedler,Maria R. Farcet,Martin Purtscher,Thomas R. Kreil
出处
期刊:Biologicals [Elsevier]
卷期号:83: 101693-101693 被引量:1
标识
DOI:10.1016/j.biologicals.2023.101693
摘要

Each process step in the manufacture of biological products requires expensive resources and reduces total process productivity. Since downstream processing of biologicals is the main cost driver, process intensification is a persistent topic during the entire product life cycle. We present here one approach for the intensification of bioprocesses by applying on-column virus inactivation using solvent/detergent (S/D) treatment during ion-exchange chromatography. The established purification process of a recombinant protein was used as a model to compare key process parameters (i.e., product yield, specific activity, impurity clearance) of the novel approach to the standard process protocol. Additional wash and incubation steps with and without S/D-containing buffers were introduced to ensure sufficient contact time to effectively eliminate enveloped viruses and to significantly decrease the amount of S/D reagents. Comparison of key process parameters demonstrated equivalent process performance. To assess the viral clearance capacity of the novel approach, XMuLV was spiked as model virus to the chromatographic load and all resulting fractions were analyzed by TCID50 and RT-qPCR. Data indicates the inactivation capability of on-column virus inactivation even at 10% of the nominal S/D concentration, although the mechanism of viral clearance needs further investigation.
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