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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 18, No. 4, 2015, pp. 302-306
Bioline Code: ej15049
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 18, No. 4, 2015, pp. 302-306

 en Expression and purification of soluble single-chain Fv against human fibroblast growth factor receptor 3 fused with Sumo tag in Escherichia coli check for this species in other resources
Liu, Zixuan; Zhang, Jizhou; Fan, Hongqiong; Yin, Ruofeng; Zheng, Zhong; Xu, Qian; Liu, Qing; He, Haiting; Peng, Xiaofan; Wang, XinXin; Li, Xiaokun & Xiao, Yechen

Abstract

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-β-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 released from the fusion protein, and further purified by Ni-NTA chromatography. The purity of ScFv was shown to be higher than 95% and their yield reached 4 mg per liter of bacterial culture. In vitro data showed that ScFv can significantly attenuate FGF9-induced phosphorylation of FGFR3.
Conclusion: We provide a novel method to produce soluble expression and bioactive functions of ScFv in Escherichia coli check for this species in other resources .

Keywords
Escherichia coli; Fibroblast growth factor receptor 3, Single-chain; Fv antibody; Soluble expression; Sumo tag

 
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