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. |
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