Prokaryotic Expression of Human β-actin Protein and Preparation of Its Monoclonal Antibody

In order to obtain hybirdoma cells secreting monoclonal antibody(McAb) directly against human β-actin,BALB/c mice were immunized with purified human β-actin protein which prokaryotic expressed from E.coli.After cell fusion and being screened,one hybridoma cell strain with the ability to stably secrete McAb against human β-actin was obtained,named as 2B4.The specificity,stability and the applicable scope of McAb were determined by indirect ELISA and western blot.The results indicated: The molecular weight of recombinant protein which could resolve in 8 mol/L urea was about 43kDa.The titers of secreting antibody from cell culture supernatant were 1×105,while 1×107 from ascites of the mice.ELISA results showed that hybridoma cells which in vitro after 20 generations and freeze-stored 3 months in liquid nitrogen could secret McAb as normal.The titer of McAb which was stored in 37 ℃ began to fall after 24 h.Western blot results showed that secreted McAb could specifically react with β-actin protein of human,mice,rabbit and fish.Hybridoma strain that secrete McAb specific against β-actin was produced.McAb will be wildly used in cell biology and immunity experiment and has a good application value.