Preparation And Identification Of Monoclonal Antibody Against D-dimer

Objective:Thrombotic diseases are very common in clinical disease, and threat to human health seriously; its morbidity and mortality are on top of various diseases, still increasing in recent years and become younger in average age. It has become one of research priorities and hot spots in the modern medicine industry. Produce the anti-D-dimer monoclonal antibody has many advantages, such as high prices, high purity, strong specificity, reproducibility, no cross-reaction and unlimited number; it can make early diagnosis of thrombosis and antibody-targeted thrombus imaging more rapid, accurate and simple.Methods:This experiment used human origin D-dimer standard as antigen immunize BALB/c female mice, get the immune spleen cells with myeloma cell SP2/0 for fusion, hybridoma cells are cultured in HAT medium; the experiment used D-dimer standard as antigen screen hybridoma which secrete anti-D-dimer antibody in indirect ELISA test, used cloning by limiting dilution to get positive hybridoma cell strain which secrete anti-D-dimer monoclonal antibody stably; this experiment also use ELISA, Western blotting testing ect. to identify feature of anti-D-dimer monoclonal antibody.Results:This experiment screened two hybridoma cell strains which secrete anti-D-dimer monoclonal antibody stably, named 6A3 and 8A9. The two monoclonal antibodies immunoglobulin subclasses were IgG2a. By ELISA, Western blotting and other methods of identification, the anti-D-dimer monoclonal antibody can specific specifically react with D-dimer.Conclusions:This experiment successfully produced the monoclonal antibody with strong affinity, high-titer, and can specifically recognize D-dimer, it will pay an important role in diagnosis and treatment of thrombosis.