Gene Construction, Expression And Internalization Analysis Of Human Anti-HBsAg Single Chain Antibody/EGFP Fusion Proteins Containing Arg9

Objective: To construct, express and purify ScFv14/EGFP fusion proteins which containing Arg9, and to study their binding activities and internalization abilities.Method: A series of oligonucleotide primers were designed to recombined Arg9 into 5'terminal of ScFv14 gene, 5'terminal or 3'terminal of EGFP gene, and used for PCR. Gene of R9/ScFv14 or ScFv14 were ligated with R9/EGFP,EGFP/R9 or EGFP gene respectively before they were cloned into the expression vector pET32a. After being induced in E.coli.BLH by IPTG and purified by Ni-NTA, the binding activities and internalization abilities of purified products were analyzed by indirect ELISA and indirect immunofluorescent analysis respectively.