使用实验设计方法优化抗CD22单链抗体片段-凋亡素融合蛋白的表达
Expression Optimization of Anti-CD22 scFv-Apoptin Fusion Protein Using Experimental Design Methodology.
作者信息
Agha Amiri Solmaz, Zarei Najmeh, Enayati Somayeh, Azizi Mohammad, Khalaj Vahid, Shahhosseini Soraya
机构信息
Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Medical Biotechnolgy, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
出版信息
Iran Biomed J. 2018 Jan 1;22(1):66-9. doi: 10.22034/ibj.22.1.66. Epub 2017 Jul 10.
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.
背景
实验设计是优化重组蛋白生产过程的一种快速且经济高效的方法。在我们之前的研究中，我们构建了一种强效双功能融合蛋白，抗CD22单链抗体片段-凋亡素，用于靶向B细胞恶性细胞系。在本研究中，我们报告了不同变量对该融合蛋白表达水平的影响。
方法
使用实验设计测试了四个变量（诱导时的细胞光密度、异丙基-β-D-硫代半乳糖苷（IPTG）浓度、诱导温度和诱导时间）。
结果
我们的研究结果表明，在所检测的变量中，只有诱导时间对蛋白表达产量有显著的正向影响。
结论
实验设计在本研究中得到成功应用。本研究中获得的优化条件可应用于该新型融合蛋白未来的商业化生产。
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