Chemiluminescence Immunoassay Imaging

This thesis consists of two parts. The first part is review, and the second one is research reports.The review includes the history of immunoassay, the basic principles, characteristics and system of chemiluminescence immunoassay (CLIA), and the immunoassay labeled technologies. Moreover, the applications of CLIA in recent years and coupling technologies in CLIA are reviewed. CLIA that uses a CL species or an enzyme as a label to study the reaction of antigen and antibody, we can determinate the CL intensity by affiliating the oxidant or enzyme to the reaction after immunoassay reaction. In this way, the concentration of sample can be gained according to determinating the intensity. The main advantages of CLIA are that they are high sensitivity, wide dynamic range, and long activity of the label, non-radiation and automatic control. With the development of new analytical methods such as flow injection, HPLC, capillary electrophoresis, and CCD camera imaging, CLIA has more application in practical analysis. The current developments of CLIA are that they should synthesize new labels in immunoassay analysis, and make as perfect or effective as possible in the immunoassay labeled technologies.The research part includes four sections, the former three sections of which are based on double antibody sandwich assay format of CLIA. In the first section, a pair of antibodies that recognize different epitopes of same antigen were used to capture and detect a certain antigen. The assay protocols were the following: (a) the 96-well plates were coated with (3-human chorionic gonadotropin monoclonal antibody. All further steps were performed at 37° C. These wells were washed three times with PBST solution; (b) addition of the P-HCG standard or sample incubation for 30 minutes, washed four times; (c) addition of HRP-conjugated anti-p-HCG monoclonal antibody. After the final washing step, the chemiluminescent substrate was added. Chemiluminescence imaging measurements were performed to detecte p-HCG with the Luminol/H2O2 /HRP/enhanced system in urine. The linear range of P-HCG was 12.5 to 400 mIU/mL and the detection limit was 4 mIU/mL. P-HCG contents in women urine were determined and the analysis results agreed well with those from hospital.