RESUMEN
AIM: To obtain purificated his-XCL1 recombinant protein of porcine in vitro and prepare the associated polyclonal antibody; To study how the recombinant protein affects on lymphocytes proliferation. METHODS: Purify the recombinant protein using HiTrap(TM); Chelating HP chromatographic column; Check the purified product using SDS-PAGE; Detect the expression of the recombinant protein using Western blot; Immunize the experimental animals with the purified fusion protein to prepare the serum containing the associated polyclonal antibody. The serum will then undergo double immnodiffusion test and indirect ELISA test to determine the polyclonal antibody titer. Then, test the condition of lymphocytes proliferation by MTT. RESULTS: Single target strip could be seen under conducting the SDS-PAGE electrophoresis when the concentration of the binding buffer is 40 mmol/L and the concentration of the elution buffer is 500 mmol/L; Western blot test showed that the recombinant protein could be successfully expressed; Double immunodiffusion test showed the antigen-antibody binding ratio to be 1:8, and the titre of antibodies was 1:12 800 detected by indirect ELISA. The result of MTT showed that both the native XCL1 and the recombinant protein could stimulate lymphocytes proliferation, and this stimulating effect could be effectively blocked by the polyclonal antibody we prepared. CONCLUSION: To conclude, this recombinant protein has biological activity and this research can provide basic material for further investigation of the function of XCL1 in swine.
