Purification and Refolding of Overexpressed Human Basic Fibroblast Growth Factor in Escherichia coli.

Abstract : This work describes the integration of expanded bed adsorption (EBA) and adsorptive protein refolding operations used to recover purified and biologically active human basic fibroblast growth factor from inclusion bodies expressed in E. coli. Insoluble overexpressed human basic fibroblast growth factor has been purified on CM Hyper Z matrix by expanded bed adsorption after isolation and solubilization in 8 M urea. The adsorption was made in expanded bed without clarification steps such as centrifugation. Column refolding was done by elimination of urea and elution with NaCl. The human basic fibroblast growth factor was obtained as a highly purified soluble monomer form with similar behavior in circular dichroism and fluorescence spectroscopy as native protein. A total of 92.52% of the available human basic fibroblast growth factor was recovered as biologically active and purified protein using the mentioned purification and refolding process. This resulted in the first procedure describing high-throughput purification and refolding of human basic fibroblast growth factor in one step and is likely to have the greatest benefit for proteins that tend to aggregate when refolded by dilution.
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Mona Alibolandi, Hasan Mirzahoseini. Purification and Refolding of Overexpressed Human Basic Fibroblast Growth Factor in Escherichia coli.. Biotechnol Res Int, 2011, 2011, pp.973741. ⟨10.4061/2011/973741⟩. ⟨pasteur-00749213⟩

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