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从转基因水稻种子中提取和纯化人白细胞介素-10。

Extraction and purification of human interleukin-10 from transgenic rice seeds.

作者信息

Fujiwara Yoshihiro, Aiki Yasuhiko, Yang Lijun, Takaiwa Fumio, Kosaka Akemi, Tsuji Noriko M, Shiraki Kentaro, Sekikawa Kenji

机构信息

Preventec, Inc., 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan.

出版信息

Protein Expr Purif. 2010 Jul;72(1):125-30. doi: 10.1016/j.pep.2010.02.008. Epub 2010 Feb 14.

Abstract

Recombinant protein production system using transgenic rice grain offers many advantages in higher accumulation, preservation, lower production cost, ease of scale up and low risk of contamination by toxic materials. We developed a transgenic rice strain whose seeds accumulate human interleukin (IL)-10, a cytokine that suppresses inflammation-related immune responses. We also developed a method of extracting and purifying IL-10 from rice seeds. A biochemical crosslinking method was used to detect the biologically active noncovalent dimer of IL-10. This method was useful for developing efficient methods of refolding and purification. The purified IL-10 comprised only noncovalent dimers and showed higher activity than the commercial IL-10. The purified IL-10 had very low endotoxin contamination and is expected to have broad clinical application.

摘要

利用转基因水稻种子的重组蛋白生产系统在更高的积累量、保存、更低的生产成本、易于扩大规模以及低毒性物质污染风险等方面具有诸多优势。我们培育了一种转基因水稻品系,其种子积累人白细胞介素(IL)-10,这是一种抑制炎症相关免疫反应的细胞因子。我们还开发了一种从水稻种子中提取和纯化IL-10的方法。采用生化交联方法检测IL-10具有生物活性的非共价二聚体。该方法有助于开发高效的重折叠和纯化方法。纯化后的IL-10仅由非共价二聚体组成,且比市售IL-10具有更高的活性。纯化后的IL-10内毒素污染极低,有望具有广泛的临床应用前景。

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