Immunoassay Protocols Selected and Optimized for 19-Nortestosterone

Objective:19-Nortestosterone (NT) has been forbidden in sports and animal racing,also been banned in food producing animals to accelerate weight gain and improve feeding efficiency.This article disscussed the rapid immunoassay selection for 19-NT.Method:Indirect ELISA was employed to evaluate the effects of immunity time on titers,homologous and heterologous indirect competitive ELISA to judge the effects on sensitivity,different incubation time to select the optimized immunoassay protocol.Result:The heterologous ELISA immunoassay with the NT pAb from rabbits immunized with NT-17-BSA as artificial antigen and NT-3-OVA as coating antigen,showed the highest titer (1:25600) and the most sensitive IC50 value (18ng/mL).At the same time,Satisfied absorbance values were obtained when the incubation protocol with 2 hours of coating time and 1 h of blocking time at 37℃ was considered.Conclusion:The established immunoassay could be used to develop rapid test kits for NT detection.