Kim Seo Woo, Lee Jeong Hyo, Han Ji Seon, Shin Seung Pyo, Park Tae Sub
Graduate School of International Agricultural Technology, Seoul National University, Pyeongchang-gun 25354, Gangwon-do, Korea.
Institute of Green-Bio Science and Technology, Seoul National University, Pyeongchang-gun 25354, Gangwon-do, Korea.
Animals (Basel). 2021 May 26;11(6):1554. doi: 10.3390/ani11061554.
A bioreactor can be used for mass production of therapeutic proteins and other bioactive substances. Although various methods have been developed using microorganisms and animal cells, advanced strategies are needed for the efficient production of biofunctional proteins. In microorganisms, post-translational glycosylation and modification are not performed properly, while animal cell systems require more time and expense. To overcome these problems, new methods using products from transgenic animals have been considered, such as genetically modified cow's milk and hen's eggs. In this study, based on a non-viral transposition system, we generated transgenic bioreactor chickens that produced human cystatin C (hCST3). There were no differences in the phenotype or histochemical structure of the wild-type and hCST3-expressing transgenic chickens. Subsequently, we analyzed the hCST3 expression in transgenic chickens, mainly in muscle and egg white, which could be major deposition warehouses for hCST3 protein. In both muscle and egg white, we detected high hCST3 expression by ELISA and Western blotting. hCST3 proteins were efficiently purified from muscle and egg white of transgenic chickens using a His-tag purification system. These data show that transgenic chickens can be efficiently used as a bioreactor for the mass production of bioactive materials.
生物反应器可用于大规模生产治疗性蛋白质和其他生物活性物质。尽管已经开发出了利用微生物和动物细胞的各种方法,但高效生产生物功能蛋白仍需要先进的策略。在微生物中,翻译后糖基化和修饰不能正常进行,而动物细胞系统则需要更多的时间和费用。为了克服这些问题,人们考虑了使用转基因动物产品的新方法,如转基因牛奶和鸡蛋。在本研究中,基于非病毒转座系统,我们培育出了能产生人胱抑素C(hCST3)的转基因生物反应器鸡。野生型和表达hCST3的转基因鸡在表型或组织化学结构上没有差异。随后,我们分析了转基因鸡中hCST3的表达情况,主要是在肌肉和蛋清中,这可能是hCST3蛋白的主要沉积库。在肌肉和蛋清中,我们通过酶联免疫吸附测定(ELISA)和蛋白质免疫印迹法检测到了高表达的hCST3。使用His标签纯化系统从转基因鸡的肌肉和蛋清中高效纯化出了hCST3蛋白。这些数据表明,转基因鸡可有效地用作生物反应器,用于大规模生产生物活性物质。