Considerations on the flow configuration of membrane chromatography devices for the purification of human basic fibroblast growth factor from crude lysates

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• 作者：Gesa Gieseler ; Iliyana Pepelanova ; Annette Meyer ; Louis Villain ; Sascha Beutel ; Ursula Rinas and Thomas Scheper
• 刊名：Engineering in Life Sciences
• 出版年：2016
• 出版时间：November 2016
• 年：2016
• 卷：16
• 期：8
• 页码：697-705
• 全文大小：1112K
• ISSN：1618-2863

文摘

Membrane chromatography possesses numerous advantages such as operation at high flow rates, low back pressure, ease of handling and scale up, which make the membrane adsorber process a viable alternative to conventional packed column chromatography. A purification process for the isolation of human recombinant basic fibroblast growth factor (FGF-2) based on membrane chromatography was investigated using devices with different flow configurations. In the first process, the FGF-2 capture step was performed with an axial flow device, while the alternative method achieved direct capture of FGF-2 from unclarified cell lysate with a tangential flow device. In both processes, FGF-2 purities exceeded 82% and the purified cytokine displayed high biological activity. Binding capacity (BC) from fermentation broth of the axial flow device was 28 mg/mL. This was 50% higher than the BC obtained with the tangential flow device under particle-free supernatant conditions (18 mg/mL) and 150% higher compared to the BC achieved with unclarified cell lysate (11 mg/mL). While membrane chromatography in tangential flow mode omits clarification and thus reduces the number of stages in the downstream process, it displays lower peak resolution and leads to a lower overall process yield.