基于表面等离子体共振技术和磁性微球免疫荧光技术对肿瘤标志物的新型分析方法分析-分析化学专业论文.docxVIP

AB ABSTRACT PAGE PAGE IV ABSTRACT Compared to the traditional methods of determination of cancer marker, surface plasmon resonance (SPR) is one of the powerful analytical tools for directly monitoring molecular interactions, with free label to the target molecules. Due to some advantages, such as fast separation, high efficiency, keep high activity, immonumagnetic beads (IMBs) have been used widely in biological analysis. In this paper, we developed a novel method to detect Carcinoembryonic antigen (CEA) in buffer and human serum spiking samples using a SPR biosensor with gold nanoparticles (GNPs) to enhance signal. Different methods were used to enhance and amplify the signal including sandwich immunoassay and the second signal amplification by GNPs. A novel immunoassay for the determination of CEA was established by combining a fluoroimmunoassay and immunomagnetic separation. In the first chapter, it has been described the principles of SPR technology, tumer marker and IBMs, their advantages, instrument, the detection format and their applications. My mayor work was illustrated. This thesis mainly focuses on development of rapid, sensitive detection methods of CEA by using SPR technology and immonumagnetic beads fluoroimmunoassay. In the second chapter, a standard direct binding assay and sandwich assay were carried out to detect CEA using SPR sensor. Antibody coupling conditions were optimized, including pH, flow rate, concentration. pH 4.5, 10 μL/min, 100μg/mL were chosed as the condition of optimization in immobilaizition processes. The immobilization of antibodies on the sensor chip caused a shift of 8475 RU, which corresponds to 8.475 ng mm-2. The the limit of detection (LOD) is 13.78 ng/mL in direc format assay. To improve the sensitivity, sandwich direct format was used to detect CEA. The LOD of this enhanced sandwich assay for CEA detection was reduced to be 3.30 ng/mL. The method of the sandwich assay with mAbCEA-B5 gave about 4.2-fold enhancement in t