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The Preparation, Purification of CEA MAB and Establishment of Double Antibodies Sandwich ELISA

Author: XieYongHua
Tutor: ZhangYanDing
School: Fujian Normal University
Course: Developmental Biology
Keywords: CEA Monoclonal antibody Protein - G affinity chromatography. Double-antibody sandwich ELISA Reaction conditions
CLC: R446.6
Type: Master's thesis
Year: 2010
Downloads: 69
Quote: 0
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Abstract


Carcinoembryonic antigen (CEA) is a cell adhesion -related glycoprotein . Colorectal cancer , gastric cancer , pancreatic cancer , lung cancer , breast cancer and medullary thyroid cancer in patients with serum CEA levels were significantly higher than in normal individuals . Carcinoembryonic antigen is used as a broad spectrum of tumor markers applied to determine the efficacy of colorectal cancer, breast cancer , lung cancer and other cancers , the progression of the disease , monitoring and prognosis . Monoclonal antibody is secreted by a single immune cells after cloning refers to a class of antibody against an antigenic epitope . To act as a substrate can produce a monoclonal antibody binding on the kind of substrate on its specificity , the monoclonal antibodies can be used to detect and identify the corresponding substrate . The ELISA method is the efficient catalysis of the specificity of the antigen-antibody reaction and enzyme combination , create a non-radioactive labeling immunoassay technology . Such technology can effectively detect the antigen or antibody in the sample content . Compared with other antibody labeling reaction method , ELISA has many advantages, such as high specificity , high sensitivity, inexpensive and safe and reliable . Carcinoembryonic antigen monoclonal antibody ELISA technique for the detection of cancer clinical applications of practical significance . The issues existing in our laboratory CEA monoclonal antibody cell line production qualified CEA monoclonal antibody with similar foreign products with the same potency . Established and optimized double-antibody sandwich ELISA method , in order to facilitate the clinical detection of CEA . The main research work involves: ( 1 ) hybridoma cells using the limiting dilution method , cell lines with high expression efficiency filter ; the expression efficiency of the cell lines after screening approximately two-fold increase than the original . ( 2 ) the use the mouse ascites production of monoclonal antibody ; (3 ) CEA monoclonal antibody ascites was purified using protein - G affinity chromatography step purification method ; ( 4 ) for the production of CEA monoclonal antibody HRP mark and its potency ; (5) double-antibody sandwich ELISA method for detection of CEA to establish the optimum reaction conditions .

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