The Molecular Cloning,Expression of Recombinant Human IL-1β and the Preparation of Its Monoclonal Antibody

Objective To clone recombinant human IL-1B gene and express with a prokaryotic expression system and to prepare the monoclonal antibody against it.Methods The cDNA of HL-60 cell line was used as a template,the human IL-1β gene was detected by PCR.IL-1β expressed by a prokaryotic expression system was purified.Mice were immunized with purified fusion protein IL-1β for three times.The specificity and sensitivity of anti-human IL-1β monoclonal antibody were characterized by Western blot and indirect ELISA.Results Fusion protein IL-1β was highly expressed in E.coli with a molecular weight of about 51kd,its purity was 95%.Western blot and FACS results showed that the monoclonal antibodies could specifically recognize the target protein expressed in the E.coli expression system and HL-60 cell line.Conclusions The IL-1βsene is successfully cloned,the fusion protein of which and the mAb against recombinant human IL-1β are prepared,which provids a useful tool for laboratory and clinical research.