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Expression and purification of soluble single-chain Fv against human fibroblast growth factor receptor 3 fused with Sumo tag in Escherichia coli Electron. J. Biotechnol.
Liu,Zixuan; Zhang,Jizhou; Fan,Hongqiong; Yin,Ruofeng; Zheng,Zhong; Xu,Qian; Liu,Qing; He,Haiting; Peng,Xiaofan; Wang,XinXin; Li,Xiaokun; Xiao,Yechen.
Background Overexpression or mutated activation of Fibroblast growth factor receptor 3 (FGFR3) is involved in the pathogenesis of many tumors. More and more studies focus on the potential usage of therapeutic antibodies against FGFR3. Results In this study, a novel single-chain Fv (ScFv) against FGFR3 was prepared and characterized. To achieve the soluble expression, ScFv was fused with Sumo (Small ubiquitin-related modifier) by polymerase chain reaction (PCR), and cloned into pET-20b. The recombinant bacteria were induced by 0.5 mM Isopropyl-ß-d-thiogalactopyranoside (IPTG) for 16 h at 20°C, and the supernatant liquid of Sumo-ScFv was harvested and purified by Ni-NTA chromatography. After being cleaved by the Sumo protease, the recombinant ScFv was...
Tipo: Journal article Palavras-chave: Escherichia coli; Fibroblast growth factor receptor 3; Single-chain Fv antibody; Soluble expression; Sumo tag.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000400008
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