Abbasi-Malati Zahra, Amiri Fatemeh, Mohammadipour Mahshid, Roudkenar Mehryar Habibi
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Department of Medical Laboratory Sciences, School of Paramedicine, Hamadan University of Medical Sciences, Hamadan, Iran.
Mol Biotechnol. 2019 May;61(5):317-324. doi: 10.1007/s12033-019-00160-y.
The mammalian expression system plays a key central role in the production of therapeutic recombinant proteins. Conspicuously, any improvements in the expression system which lead to a higher expression level would have an impact especially in bio-pharmaceutical industries. In the current study, to take steps toward the improvement of expression of recombinant protein, first, we established a stable HEK293 cell line to overexpress a well-known cytoprotective and antioxidant gene, Nrf2. Next, we transiently expressed human recombinant coagulation factor VII, as an example of human recombinant protein, in the engineered-HEK293 cell line. Our results revealed that the established cells had a higher growth rate and were able to endure to UV-induced oxidative stress. Furthermore, within our expectation, our results revealed that the expression level of recombinant FVII in Nrf2-engineered HEK293 cells (315 ng/ml) was higher than the HEK293 (198 ng/ml) cells and it was functional in a coagulation test assay. Moreover, our new cell line could be a suitable cell to express other recombinant proteins especially for large-scale production of recombinant protein under other culture condition such as lower serum and suspension culture that imposed advantages especially in terms of cost benefits in bio-pharmaceutical industries.
哺乳动物表达系统在治疗性重组蛋白的生产中起着关键核心作用。显然,表达系统的任何改进若能带来更高的表达水平,将尤其对生物制药行业产生影响。在当前研究中,为了采取措施提高重组蛋白的表达,首先,我们建立了一个稳定的HEK293细胞系来过表达一个著名的细胞保护和抗氧化基因Nrf2。接下来,我们在工程化的HEK293细胞系中瞬时表达人重组凝血因子VII,作为人重组蛋白的一个例子。我们的结果表明,所建立的细胞具有更高的生长速率,并且能够耐受紫外线诱导的氧化应激。此外,不出我们所料,我们的结果表明,重组FVII在Nrf2工程化的HEK293细胞中的表达水平(315纳克/毫升)高于HEK293细胞(198纳克/毫升),并且在凝血试验中具有功能。此外,我们的新细胞系可能是表达其他重组蛋白的合适细胞,特别是在其他培养条件下,如低血清和悬浮培养,这在生物制药行业的成本效益方面具有优势,尤其适合大规模生产重组蛋白。