Granulocyte-colony stimulating factor analog production by recombinant Escherichia coli

---

Tezin Türü: Yüksek Lisans

Tezin Yürütüldüğü Kurum: Orta Doğu Teknik Üniversitesi, Mühendislik Fakültesi, Kimya Mühendisliği Bölümü, Türkiye

Tezin Onay Tarihi: 2017

Öğrenci: ONUR ERSOY

Danışman: PINAR ÇALIK

Özet:

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.