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水稻(Oryza sativa L.)重组人转铁蛋白的表达、纯化及特性分析

Expression, purification, and characterization of recombinant human transferrin from rice (Oryza sativa L.).

作者信息

Zhang Deshui, Nandi Somen, Bryan Paula, Pettit Steve, Nguyen Diane, Santos Mary Ann, Huang Ning

机构信息

Ventria Bioscience, 2860 W Covell Blvd., Suite 1, Davis, CA 95616, United States.

出版信息

Protein Expr Purif. 2010 Nov;74(1):69-79. doi: 10.1016/j.pep.2010.04.019. Epub 2010 May 4.

Abstract

Transferrin is an essential ingredient used in cell culture media due to its crucial role in regulating cellular iron uptake, transport, and utilization. It is also a promising drug carrier used to increase a drug's therapeutic index via the unique transferrin receptor-mediated endocytosis pathway. Due to the high risk of contamination with blood-borne pathogens from the use of human or animal plasma-derived transferrin, recombinant transferrin is preferred for use as a replacement for native transferrin. We expressed recombinant human transferrin in rice (Oryza sativa L.) at a high level of 1% seed dry weight (10 g/kg). The recombinant human transferrin was able to be extracted with saline buffers and then purified by a one step anion exchange chromatographic process to greater than 95% purity. The rice-derived recombinant human transferrin was shown to be not only structurally similar to the native human transferrin, but also functionally the same as native transferrin in terms of reversible iron binding and promoting cell growth and productivity. These results indicate that rice-derived recombinant human transferrin should be a safe and low cost alternative to human or animal plasma-derived transferrin for use in cell culture-based biopharmaceutical production of protein therapeutics and vaccines.

摘要

转铁蛋白是细胞培养基中使用的一种重要成分,因为它在调节细胞铁摄取、运输和利用方面起着关键作用。它也是一种有前景的药物载体,可通过独特的转铁蛋白受体介导的内吞途径提高药物的治疗指数。由于使用人或动物血浆来源的转铁蛋白存在血液传播病原体污染的高风险,重组转铁蛋白更适合用作天然转铁蛋白的替代品。我们在水稻(Oryza sativa L.)中高水平表达了重组人转铁蛋白,其含量达到种子干重的1%(10 g/kg)。重组人转铁蛋白能够用盐缓冲液提取,然后通过一步阴离子交换色谱法纯化至纯度大于95%。结果表明,水稻来源的重组人转铁蛋白不仅在结构上与天然人转铁蛋白相似,而且在可逆铁结合以及促进细胞生长和生产力方面在功能上与天然转铁蛋白相同。这些结果表明,水稻来源的重组人转铁蛋白应该是用于基于细胞培养的蛋白质治疗药物和疫苗生物制药生产中,人或动物血浆来源转铁蛋白的一种安全且低成本的替代品。

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