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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
Author(s) -
Venkiteshwaran Adith,
Heider Patrick,
Matosevic Sandro,
Bogsnes Are,
Staby Arne,
Sharfstein Susan,
Belfort Georges
Publication year - 2008
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp0700253
Subject(s) - microfiltration , diafiltration , lysis , membrane , chemistry , escherichia coli , chromatography , cell disruption , cross flow filtration , fouling , membrane fouling , inclusion bodies , downstream processing , permeation , cell membrane , chemical engineering , biochemistry , engineering , gene
Cross‐flow membrane microfiltration was used under optimal conditions to recover met‐growth hormone 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‐step process in which the goal was to isolate IBs at high yield from the HCP. These undesired soluble HCP 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 ionic strengths to determine the optimum combination for HCP removal. Diafiltration, the washing away of impurities with protein‐free buffer, was then employed to ensure removal of the host cell proteins at the optimum conditions. About 90% removal of the HCP was obtained in about 5 diavolumes, maintaining high protein transmission and low membrane fouling.