Development of rapid immunoassay for aminophylline

• 1、School of Food Science and Technology,Jiangnan University,Wuxi Jiangsu 214122

Abstract：Monoclonal antibody against aminophylline was raised and used to develop an indirect competitive enzyme-linked immunoassay(ic-ELISA). The haptenactive conjugated to bovin serum albumin(BSA) and ovalbumin(OVA) as immounogen and coating antigen via 1-Ethyl-carbodiimide hydrochloride(EDC) and dicyclohexylcarbodiimide(DCC), respectively. One hybridoma-cell lines 1G5 secreting monoclonal antibodie against aminophylline was prepared by using mergenced myeloma cells SP2/0 of mouse with spleen cells from Balb/c mice immunized by the immounogen. The effect of several physicochemical factors on the monoclonal antibody for parathion was studied and the results were used to optimize the assay system. The IC50 was 5.80 ng/ml and the linear detection range (IC20~IC80) was 1.62 ng/mL~20.66 ng/mL measured by an indirect competitive ELISA. Cross-reactivities on acefylline was less 20%. The range of recovery rates of aminophylline from feed samples was 94%~114% and the variation coefficient was less than 10%. So monitoring aminophylline by using the monoclonal antibody is capable.

Keywords： aminophylline monoclonal antibody ic-ELISA