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Your search keyword '"Ruofeng Yin"' showing total 2 results
Start Over Author "Ruofeng Yin" Publisher pontificia universidad catolica de valparaiso
2 results on '"Ruofeng Yin"'

1. Expression and purification of soluble single-chain Fv against human fibroblast growth factor receptor 3 fused with Sumo tag in Escherichia coli

Author

Zhong Zheng, Hongqiong Fan, Yechen Xiao, Qian Xu, Xiaofan Peng, Xinxin Wang, Xiaokun Li, Zixuan Liu, Haiting He, Jizhou Zhang, Ruofeng Yin, and Qing Liu

Subjects
medicine.medical_treatment, lcsh:Biotechnology, lac operon, chemical and pharmacologic phenomena, Biology, medicine.disease_cause, Applied Microbiology and Biotechnology, law.invention, Single-chain Fv antibody, law, lcsh:TP248.13-248.65, medicine, Escherichia coli, lcsh:QH301-705.5, Soluble expression, Fibroblast growth factor receptor 3, Single-chain Fv antibody, Protease, Fibroblast growth factor receptor 3, respiratory system, Molecular biology, Fusion protein, In vitro, lcsh:Biology (General), Recombinant DNA, Phosphorylation, Biotechnology, Sumo tag
Abstract

Normal 0 21 false false false ES-CL X-NONE X-NONE 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 20oC, and the supernatant liquid of Sumo-ScFv was harvested and purified by Ni-NTA chromatography. After 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 ScFv can significantly attenuate FGF9-induced the phosphorylation of FGFR3. Conclusion: We provide a novel method to produce soluble expression and bioactive functions of ScFv in Escherichia coli . /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabla normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-ansi-language:ES-CL; mso-fareast-language:EN-US;}

Published
2015

2. Expression and purification of soluble single-chain Fv against human fibroblast growth factor receptor 3 fused with Sumo tag in Escherichia coli.

Author

Zixuan Liu, Jizhou Zhang, Hongqiong Fan, Ruofeng Yin, Zhong Zheng, Qian Xu, Qing Liu, Haiting He, Xiaofan Peng, XinXin Wang, Xiaokun Li, and Yechen Xiao

Subjects
*FIBROBLAST growth factor receptors, *POLYMERASE chain reaction, *GENE expression, *THERAPEUTIC use of immunoglobulins, *PHOSPHORYLATION
Abstract

Background: Over expression 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. [ABSTRACT FROM AUTHOR]

Published
2015
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