3M Separation and Purification Sciences, Saint Paul, Minnesota.
Johns Hopkins University, Chemical and Biological Engineering, Baltimore, Maryland.
Biotechnol Prog. 2019 Nov;35(6):e2882. doi: 10.1002/btpr.2882. Epub 2019 Aug 13.
The use and impact of 3M™ Emphaze™ AEX Hybrid Purifier, a single-use, fully synthetic chromatographic product, was explored to reduce host cell DNA (HC-DNA) concentration during the primary clarification of a monoclonal antibody (mAb). An approximately 5-log reduction in HC-DNA was achieved at an Emphaze AEX Hybrid Purifier throughput of 200 L/m . The appreciable reduction in HC-DNA achieved during primary clarification enhanced Protein A chromatography performance, resulting in a sharper and narrower elution profile. In addition, a 24× improvement in host cell protein (HCP) removal and fewer impurities nonspecifically bound to the Protein A column were observed compared to those resulting from the use of depth filtration for clarification. The use of a rapid, qualitative acidification assay to facilitate HC-DNA monitoring was also investigated. This assay involves the acidification-induced precipitation of HC-DNA, enabling the easy and rapid detection of DNA breakthrough across purification media such as Emphaze AEX Hybrid Purifier by means of turbidimetric and particle size measurements.
研究了 3M™Emphaze™AEX Hybrid Purifier 的使用和影响，这是一种即用型、全合成的色谱产品，用于降低单克隆抗体（mAb）初步澄清过程中的宿主细胞 DNA（HC-DNA）浓度。在 Emphaze AEX Hybrid Purifier 的 200L/m 的通量下，HC-DNA 减少了约 5 个对数级。在初步澄清过程中实现的 HC-DNA 显著减少，提高了 Protein A 色谱性能，导致洗脱谱更尖锐、更窄。此外，与使用深度过滤进行澄清相比，观察到宿主细胞蛋白（HCP）的去除率提高了 24 倍，并且非特异性结合到 Protein A 柱上的杂质更少。还研究了使用快速、定性酸化测定法来促进 HC-DNA 监测。该测定法涉及 HC-DNA 的酸化诱导沉淀，通过浊度和粒径测量，能够轻松快速地检测到整个纯化介质（如 Emphaze AEX Hybrid Purifier）中的 DNA 泄漏。
