Expression Optimization of Anti-CD22 scFv-Apoptin Fusion Protein Using Experimental Design Methodology

Authors

  • Mohammad Azizi Department of Medical Biotechnolgy, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
  • Najmeh Zarei Department of Medical Biotechnolgy, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
  • Solmaz Agha Amiri Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Somayeh Enayati Department of Medical Biotechnolgy, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
  • Soraya Shahhosseini Department of Pharmaceutical Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Vahid Khalaj Department of Medical Biotechnolgy, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
Abstract:

Background: Design of experiments is a rapid and cost-effective approach for optimization of recombinant protein production process. In our previous study, we generated a potent dual-acting fusion protein, anti-CD22 scFv-apoptin, to target B-cell malignant cell lines. In the present investigation, we report the effect of different variables on the expression levels of this fusion protein. Methods: Four variables (cell optical density at induction, IPTG concentration, induction temperature, and induction time) were tested using experimental design. Results: Our findings demonstrated that among the examined variables, only the induction time had a significant positive effect on the protein expression yield. Conclusion: Experimental design was successfully applied in this study. The optimized condition obtained in the current study can be applied in future commercial production of this novel fusion protein.

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Journal title

volume 22  issue 1

pages  66- 69

publication date 2018-01

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