The Preparation, Purification Of Cea Mab And Establishment Of Double Antibodies Sandwich Elisa

Carcinoembryonic antigen (CEA) is a glycoprotein involved in cell adhesion.It's not usually present in the blood of healthy adults.The levels of CEA expression in the serum of colorectal cancer, stomach cancer, pancreatic cancer, lung cancer, breast cancer and medullary thyroid carcinoma patients was significantly higher than that of the normal individuals. prognosis and monitoring of colorectal cancer, breast cancer, lung cancer and other cancers.Monoclonal antibodies (mAb) are monospecific antibodies that are identical because they are produced by one type of immune cell that are all clones of a single parent cell.Given any substance, a monoclonal antibody can be created binded to that substance specifically,which can be used to detect and identify the corresponding substrate.Enzyme-linked immunosorbent assay is a non-radioactive marker immunoassay technique,based on the antigen-antibody reaction specificity and the highly catalytic efficiency of the enzyme, which can also detect the presence of an antibody or an antigen in a sample.ELISA has many similar advantages with other monoclonal antibody reaction method, such as high specificity, high sensitivity, inexpensive and safe and reliable. ELISA technology for detecting carcinoembryonic antigen has great practical significance of the detection of cancer in clinical applications.The qualified CEA monoclonal antibody production has been produced in this research, secreted from CEA monoclonal antibodies cell lines of our laboratory. And then the double-antibody sandwich ELISA method has been established and optimized for detecting CEA in clinic. These researches include:(1)The hybridoma cells lines of high expression have been selected by the limited dilution method, which the expression efficiency of has been increased three times than that of the original cells.(2)The monoclonal antibody has been produced in mouse ascites.(3)CEA monoclonal antibody has been purified by Protein-G affinity chromatography with one step. (4)CEA monoclonal antibody has been labeled with HRP and markers and their titration. (5)The optimal reaction conditions for Double-antibody sandwich ELISA have been establised for detecting CEA.