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

Agha Amiri, Solmaz; Zarei, Najmeh; Enayati, Somayeh; Azizi, Mohammad; Khalaj, Vahid; Shahhosseini, Soraya

doi:10.22034/ibj.22.1.66

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

Authors

Solmaz Agha Amiri ¹

Najmeh Zarei ²

Somayeh Enayati ²

Mohammad Azizi ²

Vahid Khalaj ²

Soraya Shahhosseini ³

¹ Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran

² Department of Medical Biotechnolgy, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

³ Department of Pharmaceutical Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran

https://doi.org/10.22034/ibj.22.1.66

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.

Keywords

Recombinant protein

Single-chain antibodies

Fusion proteins

E. coli