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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Recombinant expression of extracellular domain of mutant Epidermal Growth Factor Receptor in prokaryotic and baculovirus expression systems
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Recombinant expression of extracellular domain of mutant Epidermal Growth Factor Receptor in prokaryotic and baculovirus expression systems

机译：原核和杆状病毒表达系统中突变表皮生长因子受体细胞外结构域的重组表达

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摘要

Epidermal Growth Factor Receptor variant III (EGFRvIII) is a tumor specific antigen detected in various tumors including gliomas, breast cancer, lung cancer, head and neck squamous cell carcinoma (HNSCC). Screening of EGFRvIII targeting drug molecules can be accelerated by developing drug screening platforms using recombinantly expressed protein. Choice of expression system is one of the major factors deciding the success of recombinant expression of a protein. In our study, we have tried to express and purify the extracellular domain (ECD) of this highly unstable protein using bacterial and baculovirus expression systems to select the expression system suited for our purpose. Even though the protein was successfully expressed in prokaryotic system, purification could be done only under denaturing conditions. But in the baculovirus expression system, the protein was expressed in soluble form and could be purified under native conditions, with single step of purification. Based on our results, we conclude that insect cells are better choice over E. coli cells for expressing EGFRvIII ECD in soluble form. This study provides insights for other researchers involved in expression of similar unstable membrane proteins, on selecting the best expression system and challenges involved. (C) 2018 Elsevier B.V. All rights reserved.
机译：表皮生长因子受体变体III（EGFRVIII）是在各种肿瘤中检测到的肿瘤特异性抗原，包括胶质瘤，乳腺癌，肺癌，头部和颈部鳞状细胞癌（HNSCC）。通过使用重组表达蛋白质显影药物筛选平台，可以加速EGFRVIII靶向药物分子的筛选。表达系统的选择是决定蛋白质的重组表达成功的主要因素之一。在我们的研究中，我们尝试使用细菌和杆状病毒表达系统表达和纯化这种高度不稳定蛋白质的细胞外结构域（ECD），以选择适合我们目的的表达系统。尽管蛋白质在原核系统中成功表达，但纯化也可以仅在变性条件下进行。但在杆状病毒表达系统中，蛋白质以可溶形式表示，可以在天然条件下纯化，具有单一的纯化。基于我们的结果，我们得出结论，昆虫细胞对大肠杆菌细胞更好地选择以可溶性形式表达EGFRVIII ECD。本研究为参与类似的不稳定膜蛋白表达的其他研究人员提供了洞察，在选择最佳表达系统和所涉及的挑战方面。 （c）2018年elestvier b.v.保留所有权利。

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