Abstract：To prepare artificial antigen of 19-nortestosterone(19-NT) and identify its immunological traits, succinic anhydride was employed to synthesis 19-NT-17-succinic anhydride ester, which was proved by(+)ESI-MS spectrum and 1H-NMR spectrogram. The 17β-19-NT ester hapten was conjugated to bovine serum albumin(BSA) as immunogen by the EDC method, and to ovalbumin(OVA) as detection antigen by mixed- anhydride technology, respectively. The results of infrared spectra(IR) and UV-visible spectra indicated that the artificial antigen was synthesized successfully and the conjugation ratio of NT-BSA was 18∶1. Balb/C mice(Mus muscalus) immunized by NT-BSA were introduced to obtain NT pAb, whose antibody titer for indirect ELISA were 1∶25 600, after the concentrations were optimized by the checkerboard method. Sensitive icELISAs were selected, showing the average IC50 value of 27.3 ng/mL, the detection range of 3.3~225.0 ng/mL, and the lowest detection limits of 1.6 ng/mL, respectively. The antisera cross-reacted with epi-nortestosterone was about 69%, but not higher than 0.05% to other hormones. The produced antibody show high sensitivity and excellent specificity, lay a potential foundation for preparing NT-Kit and rapid test strip.
