Preparation Of Monoclonal Antibodies And Development Of Immunochromatographic Strips For Fluoranthene

Polycyclic aromatic hydrocarbons(PAHs) is a kind of persistent organic pollutants with toxicities such as heritage effects, carcinogenicity and light-induced toxic effects. These compounds which are mainly origin from chemical fuel, industrial sewage and natural fires can widely distribute in water, air and soil medium making a serious threat to human health. Currently, the main detection methods for PAHs include gas chromatography, biological assay, high performance liquid chromatography and immunological detection. In recent years, many researchers pay a great attention to the immunological detection method of PAHs with its advantage of simple operation and low cost.In this study, the target substance-fluoranthene(FL) is a kind of PHAs. The complete antigens of FL were synthesized by 3- carboxyl-FL and BSA or OVA using glutaraldehyde as a bridge, which is used as immune antigen and detection antigen, respectively. Then the BALB/c mice were immunized with the FL-BSA. A cell line named A11 producting Mc Ab was achieved through cell fusion, cell selection and cell cloning. Then the ascites was obtained by injecting A11 cells into the BALB/c mice. The ascites were purified by affinity chromatography. The indirect competitive ELISA(ic-ELISA) and Immunochromatographic methods were established based on the purified Mc Ab. The conditions of the ELISA for FL detetion were optimized. The regression equation was y=-0.5578x+2.1689 with correlation coefficient R2=0.9946 and the IC50 is 1ng/m L. The range of linear detection was from 0.23 ng/m L to 4.2 ng/m L. The cross reaction is lower than 1%, except for pyrene, benzo [b] fluoranthene, benzo [a] pyrene and indeno[1,2,3-cd]pyrene with 2%, 1%, 8% and 4%, respectively. The recovery rate were 102.35%、111.48%、102.84% and 101.46% for the added 10、2.5、1.3 and 0.7 ng/m L FL to water. And the average recovery rate is 104.53% from piked sample. The concentration of FL from Daqing sample and Nanhu Park were >4.2 ng/m L and 3.44ng/m L. While FL was not detected in other samples such as Jingyue Park, Jinzhou river and potable water of Changchun.Immunochromatographic strips were prepared and the experiment conditions of different part of the strips were optimized. Finally, the concentration of test line elimination was 125 ng/m L and the concentration of the lowest detection by naked eyes was 3.8 ng/m L.No cross reaction exist, except for pyrene, benzo [a] pyrene and indeno[1,2,3-cd]pyrene with concentration higher than 100,100 and 50 ng/m L, respectively.