Pesticide Polyclonal Antibody Preparation Of2,4-D And Immune Rapid Detection Technology Of Residual Research

China is not only a large agricultural country, but also the major powers of theproduction and consumption of pesticides.2,4-D is mainly used for cereal crops,especially in wheat fields and corn fields to control annual and perennial sedges anddicotyledonous weeds. Because of its wide application, the ground water and drinkingwater have been polluted.2,4-D and its derivative commonly found in human tissue,water and soil. The traditional way to detect2,4-D is using gas phase chromatography,which needs liquid-liquid extraction, acidification separation, esterification andpurification before detection. Not only does this way need long time and vast expense,but also makes operator easily contact with some organic solvents and toxicchemicals.In this thesis, mixed anhydride method was used in the covalent bond connectionof haptene2,4-dichlorophenoxyacetic acid with albumin bovine(BSA) and eggalbumin（OVA）respectively.2,4-D-BSA was used as immunogen for New Zealandwhite rabbits to get2,4-D polyclonal antibody.Corresponding2,4-D-OVA was used in the peridium of96-well microtiter platesto detect antibody. The antibody whose titer was1：12000was produced with thesynthetical immunogen. Then indirect ELISA was established, from which IC50wasascertain as50ng/ml. The best condition for the connection of2,4-D colloidal goldprobe was optimized as well. The conjugation reaction was found to be more effectivewith the use of20nm colloidal gold particles and20ul antibody per milliliter. Whenthe added amount of potassium carbonate was15ml, the best pH condition was got.The best amount of confining liquid was60μL with the centrifugation speed of10000r/min. In the best condition, the colloidal gold test strip will work in1-2min. Inthis way, the method whose detection limit was10.0ng/mL for rapid detection of2,4-D was got.Magnetic probes labeled with different2,4-D antibodies and fluorescent probeslabeled with2,4-D-OVA were produced. After comparison, the best condition formagnetic probe was ascertained. With the standard curve, the detection limit of 260mg/mL was got.On the basic of this, the magnetic microspheres werechanged to gold magnetic particles. The gold magnetic particles found to be moreeffective with the pH of8.2and antibody of350.21g/mg in30min. Also, the conditionfor the conjugation of quantum dots fluorescent probe with the2,4-D-OVA wasoptimized. The best condition was found as the amount of the2,4-D-OVA was600μL/mL and the time for peridium was70min. In this way, the system for immunegoldmag and quantum dots fluorescence probe was produced. Through the standardcurve, the detection limit of30.907ng/mL was got. With the use of2,4-D goldmagimmune microspheres and the colloidal gold test strip principle, the goldmag test stripwas produced.