Selection and identification of full human scFv against TSLP

VernacularTitle:抗T SL P全人源单链抗体筛选与初步鉴定
Author: Jianguang ZHU ; Qing YUAN ; Li HUANG ; Wenfeng XU ; Siji NIAN
Publication Type:Journal Article
Keywords: TSLP; Human scFv; Selection
From: Chinese Journal of Immunology 2014;(12):1662-1665,1669
CountryChina
Language:Chinese
Abstract: Objective:Expression of protein TSLP and selection of full human anti-TSLP single chain Fv ( scFv).Methods:The cDNA of TSLP was amplified.The amplified target gene and the expression vector pET 101/D-TOPO were ligated , and then transformed into E.coli BL21.The protein was induced to expression by IPTG and purified and identified.The biotinylated TSLP protein was used as antigen to select of human TSLP scFv from a constructed human scFv library by phage display .Results: The size of amplified cDNA of TSLP was about 423 bp,and that of expressed protein was about 26 kD.Dot blot and Western blot results showed that the expressed protein was correct.The constructed human scFv library was enriched for three rounds using biotinylated TSLP as antigen by phage display.ELISA results showed that 35% scFvs had binding activity with TSLP.The scFvs with good binding activity were selected and identified by Western blot and sequencing.Conclusion: The full human scFvs against for TSLP were selected suc-cessfully.