New Expression Method and Characterization of Recombinant Human Granulocyte Colony Stimulating Factor in a Stable Protein Formulation

Authors

  • Ralitza Boubeva School of Pharmaceutical Sciences University of Geneva, University of Lausanne 30 Quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland
  • Christian Reichert School of Pharmaceutical Sciences University of Geneva, University of Lausanne 30 Quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland
  • René Handrick University of Applied Sciences 35 Hubertus-Liebrecht Straße, D-88400 Biberach, Germany
  • Claudia Müller School of Pharmaceutical Sciences University of Geneva, University of Lausanne 30 Quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland
  • Jürgen Hannemann School of Pharmaceutical Sciences University of Geneva, University of Lausanne 30 Quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland
  • Gerrit Borcharda School of Pharmaceutical Sciences University of Geneva, University of Lausanne 30 Quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland. Gerrit.Borchard@unige.ch

DOI:

https://doi.org/10.2533/chimia.2012.281

Keywords:

Autoinduction media, Protein expression, Protein refolding, Protein stability, Rhg-csf

Abstract

Human recombinant granulocyte colony stimulating factor (rhG-CSF) is widely used in hematology and oncology for the treatment of neutropenia, for the restoration of neutrophil production after bone marrow transplantation, for myelodysplastic syndromes, and aplastic anemia. The E. coli expression system is commonly used for fast recombinant production of rhG-CSF at a large scale. We have applied a novel autoinduction method for the batch expression of rhG-CSF to study whether this new system would increase cell mass and target-protein yield compared to conventional E. coli cell culture and induction with isopropyl ?-D-thiogalactopyranoside (IPTG). We could demonstrate 3-fold higher culture densities and a 5-fold higher protein yield compared to IPTG induction without the need to monitor cell growth in a shortened 24 h expression procedure. rhG-CSF expressed in autoinduction media was successfully extracted from E. coli inclusion bodies and refolded by dialysis. After size exclusion chromatography (SEC) purification, rhG-CSF showed similar conformation, biological activity and aggregation profile compared to the commercially available biosimilar TEVAgrastim® (TEVA Pharma AG). Expression by autoinduction is suggested as a cost- and time-effective method for rhG-CSF production.

Downloads

Published

2012-05-30

How to Cite

[1]
R. Boubeva, C. Reichert, R. Handrick, C. Müller, J. Hannemann, G. Borcharda, Chimia 2012, 66, 281, DOI: 10.2533/chimia.2012.281.

Issue

Section

Scientific Articles