Preparation Of Antibodies Against Coplanar Polychlorinated Biphenyls (Co-PCBs) And Its Application In Surface Plasmon Resonance (SPR)

Polychlorinated biphenyls (PCBs) are man-made compounds and persistent organic pollutants, and they have become one of the key research and monitoring organic pollutant in the word because they pose a potential threat to ecological environment and human. PCB126 which belongs Coplanar polychlorinated biphenyls (Co-PCBs), has the most significant effect on ecological environment and human health for its highly reactivity,strong carcinogenic,mutagenic and teratogenic effects,serious harm. So we chose PCB126 as the research object.At present, relative to the ultra-trace levels of PCBs in environment (usually ng level or pg level), the existing PCBs analysis methods (chromatography, biological and immunological methods) can not be used for the actual test, because their detection limit for PCBs is onlyμg level. Therefore, the research and development of sensitive,reliable and easy detection of PCBs analysis method is of great significance.Based these, we design the research program as follows:The first step, the gene sequence of recombinant antibody against Co-PCBs indexed was constructed, and expressed it in Escherichia coli M15, then the recombinant antibody against Co-PCB was obtained by purifying and renaturation;The second step, the Self-assembled Monolayer (SAM) and Surface Plasmon Resonance (SPR) were used to detect the specificity of antibody and the interaction between antibody and PCB126, and verify the results.In these studies, we obtained the following results:1,The antibody against Co-PCBs was obtained successfully through genetic engineering;2,The antibody against Co-PCBs was assembled successfully on the surface of gold film through SAM;3,The specificity of antibody was detected and the detection limit was about 5×10-4 pg/ L basing on SPR technology;4,The above results was verified successfully on SPR-2002.Our experimental results will provide a fast, simple and real-time detection methods for the development and research of Co-PCBs ultra-trace detection kit in the environment, and the establishment of detection methods will provide an important basis for the spatial analysis,ecological security risk assessment,pollution control and environmental management of PCBs in the major river systems of the country.