| dc.creator | Liu,Zixuan | |
| dc.creator | Zhang,Jizhou | |
| dc.creator | Fan,Hongqiong | |
| dc.creator | Yin,Ruofeng | |
| dc.creator | Zheng,Zhong | |
| dc.creator | Xu,Qian | |
| dc.creator | Liu,Qing | |
| dc.creator | He,Haiting | |
| dc.creator | Peng,Xiaofan | |
| dc.creator | Wang,XinXin | |
| dc.creator | Li,Xiaokun | |
| dc.creator | Xiao,Yechen | |
| dc.date | 2015-07-01 | |
| dc.date.accessioned | 2019-05-03T12:45:19Z | |
| dc.date.available | 2019-05-03T12:45:19Z | |
| dc.identifier | https://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000400008 | |
| dc.identifier.uri | http://revistaschilenas.uchile.cl/handle/2250/85493 | |
| dc.description | 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 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. | |
| dc.format | text/html | |
| dc.language | en | |
| dc.publisher | Pontificia Universidad Católica de Valparaíso | |
| dc.relation | 10.1016/j.ejbt.2015.05.006 | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.source | Electronic Journal of Biotechnology v.18 n.4 2015 | |
| dc.subject | Escherichia coli | |
| dc.subject | Fibroblast growth factor receptor 3 | |
| dc.subject | Single-chain Fv antibody | |
| dc.subject | Soluble expression | |
| dc.subject | Sumo tag | |
| dc.title | Expression and purification of soluble single-chain Fv against human fibroblast growth factor receptor 3 fused with Sumo tag in Escherichia coli | |
