Monoclonal Antibody Preparation And Rapid Immunoassay Development For Heavy Metal And MC-LR

River, soil and food were severely contaminated with the rapid development of economy in China. Heavy metal contamination and microcystin contamination induced by the eutrophication in water and food were the two major problems. Immunologic technology with the advantages of simple, fast and sensitivity was widespread used in the area of animal epidemic disease, food safty and biomedicine. In this research, monoclonal antibodies for heavy metal and microcystin were produced based on the principles of immunology. Rapid immunoassay technologies were developed for detecting the heavy metal and microcystin contamination in drinking water and cereal.Heavy metal immunogen and coating antigen were synthesized and uesed for immunization and detection. The monoclonal antibodies were produced by cell fusion technique. The affinity constant of the anti-Pb-ITCBE antibody secreted by cell line B5-D6 was about 1.02?109 L/mol, with an IC50 of 1.3 ng/m L. The affinity constant of the anti-Cd-EDTA antibody secreted by cell line H7-A11 was about 1.1?1010 L/mol, with an IC50 of 1.8 ng/mL. The affinity constant of anti-Hg-EDTA antibody secreted by cell line F6-G3 was about 1.65?109 L/mol, with an IC50 of 4.8 ng/m L. The affinity cnstant of anti-Cu-EDTA antibody secreted by cell line C5-B11 was about 1.26?109 L/mol, with an IC50 of 3.9 ng/m L.Indirect competitive ELISA was established for detection lead, cadmium, mecury and copper in drinking water and tap water. The recoveries were from 84.3% to 111.0%. The coefficient of variation of intra-assay was less than 10% and the coefficient of variation of interassay were from 6.3 % to 10.7 %.The immunochromatographic strip was assembled for detection lead, cadmium, mecury and copper in drinking water and tap water. The detection of limit were 2 ng/m L, 0.5 ng/m L, 1 ng/m L and 2.5 ng/m L and the cut-off values were 10 ng/m L, 2 ng/mL, 10 ng/mL and 10 ng/mL.Simple and fast solid-liquid extraction method was developed for pretreating the rice containing cadmium and quantitative immunochromatographic strip was established for detecting cadmium in rice. The LOD was 0.02 mg/kg and the correlation coefficient was 0.95 when compared with the ICP-MS for detection real samples.At last, a monoclonal antibody for MC-LR was successfully produced and an immunochromatographic strip was developed for detection MC-LR in tap water and lake water, with cut-off value of 1 ng/m L. Based on the antibodies produced, an ultrasensitive immunochromatographic assay was developed for the simultaneous detection of five chemicals containing Pb and MC-LR in drinking water.