中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
Large-scale purification of human granulocyte-macrophage colony-stimulating factor expressed in Bombyx mori pupae

文献类型:期刊论文

作者Chen, J; Nie, ZM; Lu, ZB; Zhu, CG; Xu, CZ; Jin, YF; Wu, XF; Zhang, YZ
刊名APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
出版日期2007
卷号141期号:1页码:149-159
关键词Bombyx mori nucleopolyhedrovirus silkworm pupae human granulocyte-macrophage colony-stimulating factor purification isoelectric precipitation (NH4)(2)SO4 fractional precipitation target protein
通讯作者Zhang, YZ (reprint author), Zhejiang Sci Tech Univ, Inst Biochem, Hangzhou 310018, Peoples R China.,yaozhou@zist.edu.cn
英文摘要Human granulocyte-rnacrophage colony-stimulating factor (hGM-CSF) acts on many different kinds of cells, including monocytes, macrophages, granulocytes, eosinophils, and multipotential stem cells. To explore further explore pharmaceutical action, we expressed hGM-CSF by the Bombyx 1,mori nucleopolyhedrovirus expression system in silkworm pupae. However, purifying recombinant proteins from silkworm pupae on a large scale has been a big challenge. To establish purification methods suitable for mass production, we tried two crude preparation methods: (NH,)(2)SO4 fractional precipitation and isoelectric precipitation with a combination of gel filtration and ion-exchange chromatography. The isoelectric precipitation method was found to be more efficient. With this method, we eventually obtained approx 11.7 mg of 95% pure product from 1000 g of infected silkworm pupae. The recovery of purified protein was greatly increased, by approx 40%, compared with the other method. The biologic activity of this protein was determined up to 9.0 x 10(6) colony-forming units/mg in the final purified product.
学科主题Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology
类目[WOS]Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]NUCLEAR POLYHEDROSIS-VIRUS ; HIGH-LEVEL EXPRESSION ; HUMAN GM-CSF ; BACULOVIRUS VECTOR ; SILKWORM LARVAE ; INTERFERON ; SYSTEM ; CELLS ; LACTOFERRIN ; PROTEINS
收录类别SCI
语种英语
WOS记录号WOS:000246459500012
版本出版稿
源URL[http://202.127.25.143/handle/331003/1505]  
专题上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式
GB/T 7714
Chen, J,Nie, ZM,Lu, ZB,et al. Large-scale purification of human granulocyte-macrophage colony-stimulating factor expressed in Bombyx mori pupae[J]. APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,2007,141(1):149-159.
APA Chen, J.,Nie, ZM.,Lu, ZB.,Zhu, CG.,Xu, CZ.,...&Zhang, YZ.(2007).Large-scale purification of human granulocyte-macrophage colony-stimulating factor expressed in Bombyx mori pupae.APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY,141(1),149-159.
MLA Chen, J,et al."Large-scale purification of human granulocyte-macrophage colony-stimulating factor expressed in Bombyx mori pupae".APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY 141.1(2007):149-159.

入库方式: OAI收割

来源:上海生物化学与细胞生物学研究所

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