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
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.
data showed that ScFv can significantly attenuate FGF9-induced phosphorylation of FGFR3.
We provide a novel method to produce soluble expression and bioactive functions of ScFv in