Optimized Removal of Soluble Host Cell Proteins for the Recovery of met-Human Growth Hormone Inclusion Bodies from Escherichia coli Cell Lysate Using Crossflow Microfiltration

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• 作者：Adith Venkiteshwaran ; Patrick Heider ; Sandro Matosevic ; Are Bogsnes ; Arne Staby ; Susan Sharfstein ; Georges Belfort
• 刊名：Biotechnology Progress
• 出版年：2007
• 出版时间：June 2007
• 年：2007
• 卷：23
• 期：3
• 页码：667 - 672
• 全文大小：74K
• 年卷期：v.23,no.3(June 2007)
• ISSN：1520-6033

文摘

Cross-flow membrane microfiltration was used under optimal conditions to recover met-growthhormone inclusion bodies (IBs) from Escherichia coli cell lysate by removal of the host-cell(bacterial) proteins (HCP) under minimal fouling conditions. This is the first step of a two-stepprocess in which the goal was to isolate IBs at high yield from the HCP. These undesired solubleHCP were removed by passing them through the membrane while retaining the insolubles,including the aggregated IBs. Experiments were conducted at constant permeate flux with flat-sheet membranes of different pore sizes and chemistry, with feeds of varying pH and ionicstrengths to determine the optimum combination for HCP removal. Diafiltration, the washingaway of impurities with protein-free buffer, was then employed to ensure removal of the hostcell proteins at the optimum conditions. About 90% removal of the HCP was obtained in about5 diavolumes, maintaining high protein transmission and low membrane fouling.