Granulocyte-colony stimulating factor analog production by recombinant escherichia coli

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2017

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Ersoy, Onur

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Granulocyte-colony stimulating factor (G-CSF) is a significant pharmaceutical biologic. It is predominantly produced using Escherichia coli in the industry. In this study, an E. coli expression strain was genetically engineered to heterologously express a His-tagged mutant G-CSF with one amino acid substitution with respect to native human protein. A fed-batch strategy for this strain was developed. Biomass concentration in 7.0 L bioreactor reached a density of 58 grams of dry cell weight per liter. Recombinant protein concentration of bioreactor culture reached 4.9 grams per liter. G-CSF C17A was purified from cell culture, and was shown to have comparable bioactivity to innovator product, in vitro. Purified G-CSF C17A was PEGylated to achieve a bioproduct with enhanced properties. A 37% conversion of G-CSF C17A to PEG-G-CSF 17A was achieved in PEGylation reaction.

Subject Keywords

Bioreactors., Granulocytes., Escherichia coli., Recombinant proteins.

URI

http://etd.lib.metu.edu.tr/upload/12621587/index.pdf
https://hdl.handle.net/11511/26749

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Graduate School of Natural and Applied Sciences, Thesis

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Citation Formats

O. Ersoy, “Granulocyte-colony stimulating factor analog production by recombinant escherichia coli,” M.S. - Master of Science, Middle East Technical University, 2017.