Ranjbari Javad, Babaeipour Valiollah, Vahidi Hossein, Moghimi Hamidreza, Mofid Mohammad Reza, Namvaran Mohammad Mehdi, Jafari Sevda
Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran. ; Department of Life Science Engineering, School of New Science and Technologies, University of Tehran, Iran.
Iran J Pharm Res. 2015 Summer;14(3):907-17.
Human insulin-like growth factor I (hIGF-I) is a kind of growth factor with clinical significance in medicine. Up to now, E. coli expression system has been widely used as a host to produce rhIGF-1 with high yields. Batch cultures as non-continuous fermentations were carried out to overproduce rhIGF-I in E. coli. The major objective of this study is over- production of recombinant human insulin-like growth factor I (rhIGF-I) through a developed process by recruiting effective factors in order to achieve the most recombinant protein. In this study we investigated the effect of culture medium, induction temperature and amount of inducer on cell growth and IGF-1 production. Taguchi design of experiments (DOE) method was used as the statistical method. Analysis of experimental data showed that maximum production of rhIGF-I was occurred in 32y culture medium at 32 °C and 0.05 Mm IPTG. Under this condition, 0.694 g/L of rhIGF-I was produced as the inclusion bodies. Following optimization of these three factors, we have also optimized the amount of glucose and induction time in 5 liter top bench bioreactor. Full factorial design of experiment method was used for these two factors as the statistical method. 10 g/L and OD600=5 were selected as the optimum point of Glucose amount and induction time, respectively. Finally, we reached to a concentration of 1.26 g/L rhIGF-1 at optimum condition.
人胰岛素样生长因子I（hIGF-I）是一种在医学上具有临床意义的生长因子。到目前为止，大肠杆菌表达系统已被广泛用作宿主来高产重组人胰岛素样生长因子-1（rhIGF-1）。进行分批培养作为非连续发酵，以在大肠杆菌中过量生产rhIGF-I。本研究的主要目的是通过开发一种方法，通过引入有效因素来过量生产重组人胰岛素样生长因子I（rhIGF-I），以获得最多的重组蛋白。在本研究中，我们研究了培养基、诱导温度和诱导剂用量对细胞生长和IGF-1产量的影响。采用田口实验设计（DOE）方法作为统计方法。实验数据分析表明，rhIGF-I的最大产量出现在32Y培养基中，温度为32°C，IPTG浓度为0.05 mM时。在此条件下，以包涵体形式产生了0.694 g/L的rhIGF-I。在优化这三个因素之后，我们还在5升台式生物反应器中优化了葡萄糖用量和诱导时间。对这两个因素采用全因子实验设计方法作为统计方法。分别选择10 g/L和OD600 = 5作为葡萄糖用量和诱导时间的最佳点。最后，在最佳条件下我们得到了1.26 g/L的rhIGF-1浓度。
