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Optimization of the downstream process for high recovery of rhG-CSF from inclusion bodies expressed in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
35640 45099 2008 10 PDF Available
Title
Optimization of the downstream process for high recovery of rhG-CSF from inclusion bodies expressed in Escherichia coli
Abstract

Recombinant human granulocyte colony-stimulating factor (rhG-CSF) expressed as cytoplasmic inclusion bodies (IBs) in Escherichia coli, was recovered with higher yield and purity by developing an optimized process involving the sequential evaluation of complete downstream processes. Triton X-100 and urea wash removed 90% of lipopolysaccharides, 22% of host DNA and 98% of host cell proteins, leading to inclusion bodies purity of 99% with 91% recovery. Removal of host impurities at solubilization step and refolding by diafiltration at high concentration of protein were identified as key parameters with significant impact on product recovery and purity. Desalting, size exclusion and formulation were performed in a single step. This optimized process recovery yielded 69% (34 mg from 1 g of wet cell mass) of purified protein with more than 99% purity. According to available literature, the purified rhG-CSF yield obtained in this study is the highest that has been reported so far.

Keywords
rhG-CSF; Escherichia coli; Inclusion bodies; Cell disruption; Solubilization; Refolding
First Page Preview
Optimization of the downstream process for high recovery of rhG-CSF from inclusion bodies expressed in Escherichia coli
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 43, Issue 5, May 2008, Pages 566–575
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering