Prokaryotic Expression of Extracellular Domain of Human EGFR and It's Immunogenity

The object is to investigate the end its immunogenity in mouse.A DNA fragment encoding extracellular domain of human EGFR was obtained from a previoused constructed plasmid.The DNA fragment was then ligated into prokaryotic expression vector,and was expressed in Escherichia Coli harboring the constructs.The recombinant protein within insoluble inclusion bodies was purified under denatured conditions by affinity chromatography,and the refolding of the recombinant protein was performed with gradient dialysis.The recombinant protein could induce immune response in mouse producing antibody to recognize the recombinant protein and EGFR expressed at A549 tumor cells by western blot analysis and flow cytometric analysis.These findings provided evidence of the renatured recombinant extracellular domain of human EGFR is immunogenity in mouse may be of important for the further exploration of the application of this protein in immunological and immunotherapy research.