Construction of single chain Fv phage display library of anti-human-CTGF

Abstract

Objective To construct single chain Fv (scFv) library of human connective tissue growth factor (CTGF) by assembling the gene from immunized mice to phagemid PAK100.Methods Mice were immunized with recombinant human CTGF (rhCTGF).Total RNA was extracted from splenocytes of the immunized mice.Complementary DNA was got by reverse transcription PCR with the extracted RNA.Then the fragment of VH (variable heavy) and VL (variable light) were obtained by PCR.And they were assembled into scFv,which was inserted into phagemids vector PAK100 through SfiIsites.The phagemids containing scFV were transferred into E.coli.XL1-Blue bacterial cells to construct the scFv library to anti-human CTGF.Results Agarose gel clectrophoresis and sequences of phagemids certified that scFv phage display library to anti-human-CTGF was established.Conclusions The establishment of scFv library of anti-human CTGF set a foundation to get anti-human CTGF scFv. Key words: Connective tissue growth factor; Phage display; Phage display library; Anti-human CTGF scFv