The Screening,Expression and Purification of Single Chain Fv(scFv) against Vascular Endothelial Growth Factor Receptor 2

To block the activity of vascular endothelial growth factor transmitted via vascular endothelial growth factor receptor 2 pathway,a high affinity antibody against vascular endothelial growth factor receptor 2 was screened from phage display antibody library.After PCR analysis,DNA sequencing,and sequence identification,the gene of selected antibody was transformed into E.coli HB2151.Soluble ScFv antibody was expressed in the recombinant E.coli HB2151 induced by 1mmol/L IPTG and further purified by Ni-IMAC.The yield was 614μg/L with purity of 95%.In conclusion,a scFv named AK404 was obtained which will potentially serve as drug candidate in anti-angiogenesis tumor therapy or tumor diagnosis.