Development of ELISA for the detection of neomycin: preparation of anti-neomycin antibodies

In order to obtain artificial antigen of neomycin, neomycin was linked to Bovine Serum albumin(BSA) and keyhole limpet hemocyanin(KLH) respectively by EDC coupling. The conjugates BSA-NEO and KLH-NEO were identified by infrared spectrum. The infrared spectra of conjugates showed the absorption peaks of both carrier protein and neomycin. These results identified that neomycin was linked to carrier protein successfully. Four New Zealand White rabbits were subcutaneously immunized with conjugates (BSA-NEO and KLH-NEO) and developed good antibody titers, the highest titer was 1.28×105.