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重组抗HER2单链可变片段(scFv-Fc)在拟南芥中的种子特异性表达及分析

Seed-specific expression and analysis of recombinant anti-HER2 single-chain variable fragment (scFv-Fc) in Arabidopsis thaliana.

作者信息

Dong Yuanyuan, Li Jian, Yao Na, Wang Dezhong, Liu Xiuming, Wang Nan, Li Xiaowei, Wang Fawei, Li Haiyan, Jiang Chao

机构信息

Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130118, China.

Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130118, China.

出版信息

Protein Expr Purif. 2017 May;133:187-192. doi: 10.1016/j.pep.2017.03.009. Epub 2017 Mar 9.

Abstract

Antibodies to human epidermal growth factor receptor 2 (HER2) are a key element of breast cancer therapy; however, they are expensive to produce and their availability is limited. A seed-specific expression system can be used to produce recombinant proteins. We report a seed-specific expression system for the manufacture of anti-HER2 ScFv-Fc in Arabidopsis thaliana, driven by the Phaseolus vulgaris β-phaseolin promoter. Recombinant anti-HER2 ScFv-Fc was successfully and specifically expressed in seeds, and identified by protein analysis. The highest protein accumulation level, with a maximum of 1.1% of total soluble protein, was observed in mature seeds. We also demonstrated the anti-tumor potency of the plant-derived antibody against SK-BR-3 cells. These results suggest that seed-expression systems could contribute to the manufacture of commercial antibodies such as anti-HER2 ScFv-Fc.

摘要

抗人表皮生长因子受体2(HER2)抗体是乳腺癌治疗的关键要素;然而,其生产成本高昂且供应有限。种子特异性表达系统可用于生产重组蛋白。我们报告了一种由菜豆β-菜豆蛋白启动子驱动的、用于在拟南芥中制造抗HER2单链抗体片段-免疫球蛋白融合蛋白(ScFv-Fc)的种子特异性表达系统。重组抗HER2 ScFv-Fc在种子中成功且特异性地表达,并通过蛋白质分析得以鉴定。在成熟种子中观察到了最高的蛋白质积累水平,最高可达总可溶性蛋白的1.1%。我们还证明了植物源抗体对SK-BR-3细胞的抗肿瘤效力。这些结果表明,种子表达系统有助于制造诸如抗HER2 ScFv-Fc等商业抗体。

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