A Novel Bifunctional Protein TNFR2-Fc-IL-1ra (TFI): Expression, Purification and its Neutralization Activity of Inflammatory Factors
文献类型：期刊论文
| 作者 | Xie, Bo1; Liu, Shan1; Wu, Shuyin2; Chang, Alan3; Jin, Weiyuan2; Guo, Zhixiang2; Ye, Shuaidong2; Hui, Mizhou1,2 |
| 刊名 | MOLECULAR BIOTECHNOLOGY
 |
| 出版日期 | 2013-06-01 |
| 卷号 | 54期号:2页码:141-147 |
| 关键词 | Bifunctional protein GC-rich vector Large scale cultivation Bioreactor Affinity Purification |
| 英文摘要 | Tumor necrosis factor receptor (TNF) and internleukin-1 (IL-1) are the most potent proinflammatory cytokines involving in autoimmune and inflammatory human diseases. Many anti-inflammatory agents have been exploited for anti-inflammation treatments by targeting cytokines including TNF and IL-1. Theoretically, simultaneously neutralizing or blocking two important inflammatory mediators may achieve a synergistic therapeutic effect. We have developed a recombinant fusion protein, TNFR2-Fc-IL-1ra (TFI), which consists of a TNF-neutralizing domain that specifically binds to TNF-alpha, an IL-1 receptor antagonist domain, and a dimerization Fc portion of human IgG1, for bifunctional inflammatory inhibitor. Recombinant DNA expressing the sequence of this fusion protein was expressed in CHO-S cells. The protein product was purified using a two-step purification protocol and the identity of the protein was confirmed by western blot analysis. The purified recombinant protein had a purity of about 98 % as determined by HPLC, and a molecular mass of 164.6 kDa as determined by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The results of cell binding inhibition indicate that TFI was able to strongly neutralize TNF activity and antagonize IL-1r activity, suggesting that TFI may be used as a bifunctional ligand with enhanced anti-inflammatory effect. The result obtained in this study may provide a platform for extending bifunctional anti-inflammatory drug development. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 关键词[WOS] | INTERLEUKIN-1 RECEPTOR ANTAGONIST ; RHEUMATOID-ARTHRITIS ; THERAPEUTIC TARGETS ; DISEASES ; MECHANISMS ; CYTOKINES ; ALPHA ; DNA |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000318308400003 |
| 公开日期 | 2015-05-27 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/13458]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 作者单位 | 1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 2.AmProt China Inc, Hangzhou 310019, Zhejiang, Peoples R China 3.Dalian Univ Technol, Sch Life Sci & Biotechnol, Dalian 116024, Peoples R China |
| 推荐引用方式 GB/T 7714 | Xie, Bo,Liu, Shan,Wu, Shuyin,et al. A Novel Bifunctional Protein TNFR2-Fc-IL-1ra (TFI): Expression, Purification and its Neutralization Activity of Inflammatory Factors[J]. MOLECULAR BIOTECHNOLOGY,2013,54(2):141-147. |
| APA | Xie, Bo.,Liu, Shan.,Wu, Shuyin.,Chang, Alan.,Jin, Weiyuan.,...&Hui, Mizhou.(2013).A Novel Bifunctional Protein TNFR2-Fc-IL-1ra (TFI): Expression, Purification and its Neutralization Activity of Inflammatory Factors.MOLECULAR BIOTECHNOLOGY,54(2),141-147. |
| MLA | Xie, Bo,et al."A Novel Bifunctional Protein TNFR2-Fc-IL-1ra (TFI): Expression, Purification and its Neutralization Activity of Inflammatory Factors".MOLECULAR BIOTECHNOLOGY 54.2(2013):141-147. |
入库方式： OAI收割
来源：过程工程研究所
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