| China is one of the biggest countries for production and consumption of pesticides, which results in the serious pesticide pollution. Accompanied by taboo of pesticides of high toxicity such as methamidophos, parathion and so on, chlorpyrifos, as a kind of new pesticide with low toxicity, is used widespreadly in recent years. Immunoassay, as a fast, sensitive and reliable technique, is adapted to monitoring the pesticide. In this study, we chose chlorpyrifos as target pesticide, prepared chlorpyrifos monoclonal antibody and established ELISA for chlorpyrifos.The main contents are as follows:1. Synthesis and identification of artificial antigens for chlorpyrifosMolecular weight of chlorpyrifos is only 350.62. It can not elicit an antibody response alone and must be attached to a carrier protein to form an artificial antigen first. Half antigen, O,O-diethyl-O-[3,5-dichloro-6-(2-carboxyethyl)thio-2-pyridyl] phosphorothioate was synthesized by using chlorpyrifos active compound reacted with 3-marcapropanoic acid in hot alkaline solution. Then carbodiimide (EDC) was used to conjugate half antigen to bovine serum albumin (BSA) and ovalbumin (OVA) to form immunizing antigen and coating antigen respectively.Two methods were used to identify artificial antigens. Compared with UV spectrum of carriers BSA and OVA, it was found that the maximal UV absorbance peak of artificial antigens had been changed, the maximal UV absorbance peak of the two artificial antigens had moved to far ultraviolet region which meant artificial antigens had been synthesized successfully. Phosphor concentrations of artificial antigens were determined by Phosphor Molybdate Blue Colorimeter Method, and then conjugating ratios were calculated. The results were as follows: immunizing antigen was 1∶64 and coating antigen was 1∶19.2. Preparation and screening of hybridoma cells for chlorpyrifosBalb/c mice were immunized by intraperitoneal injection of immunizing antigen. Antiserum was...
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