Construction and expression of the fusion protein consisting of IFN-? and the L chain of antibody against HBsAg / 中华传染病杂志

ABSTRACT

Objective Construct an expression vector for anti-HBs Fab fragment and IFN-? fusion protein, then transform it into E.coli Top10 in order to produce a targeting drug for hepatitis B. Methods Using PCR and molecular clone techniques, we amplify the gene fragment of IFN-? with corresponding endonuclease sites and artificial linker at 5′,3′ termini,then recombine it within the vector of anti-HBs Fab gene in correct endonuclease sites at 3′ termini of the Lc gene, choosing the positive clone with PCR and transform it into E.coli Top10,after the expression and purification of the fusion protein, we use ELISA and Dot-blot to verify the antigenicity and binding activity to HBsAg. Results We got fusion protein consisting Lc and IFN-?. It has the IFN-? activity and HBsAg affinity which is similar to anti-HBs Fab fragment. Conclusions The successful expression of the fusion protein with both HBsAg affinity and IFN-? activity make it possible for immunobiotherapy drug targeted to HBV.