| Fenvalerate is an important potential endanger in the safety of food consumption and trade and agriculture ecological environment, whereas Fenvalerate is the extensive application and residual in china currently.This reaserch select Fenvalerate, with typical chemical structure, as the research object. Designed for scheme which connecting the spacer to the hapten in a new site. Monoclonal antibody was prepared, a rapid detection method was etablished, the immune site of spacer orientation in hapten molecules affect antibody recognition was researched. To be fulled of the rapid field detection of the current requirements for fenvalerate, to provide a theoretical basis for rational design of hapten, such as pesticides and other chemical pollutants in small molecule.1. Fenvalerate, with typical chemical structure, was selected as the research object. By using organic synthesis, the type of haptens was synthesized by introducing a spacer arms in one end of the Fenvalerate, we intended to expose the acid party and the aromatic party to get the better immune effect.2. Artificial antigen was synthesized using the activated ester method. The UV spectrogram of the conjugate was found different from the hapten and carrier protein, showing that the artificial antigens were synthesized successfully. After immunization, the best titer was1:256000.3. Two cloning hybridoma cells (3A7and4G10) were screened by cell fusion. The IC50values for fenvalerate were3.548μg/L and0.5623μg/L, respectively. The limit of detection (LOD) of MAb (4G10) was0.02μg/L and less than3.7%cross-reactivity. It is showed that the best sensitivity to fenvalerate and little cross-reactivity with other pyrethroids, and explained that MAb (4G10) to fenvalerate with high sensitivity and specificity..4. After optimization, standard curve of fenvalerate was setted up. The detection range was0.02μg/L～600μg/L. Lay the foundation For future ELISA rapid detection kit and immunoaffinity micro-column prepared. |
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