Abstract: Neomycin was conjugated to carrier protein BSA and OVA by the method of EDC to form the immunizing antigen BSA2NEO and the coating antigen OVA2NEO. IR and SDS2PAGE were used to identify the NEO artificial anti2 gen.Balb/ c mice were immunized with BSA2NEO. The titre of polyclonal antibody was detected by indirect ELISA and blocking ELISA. The hybridoma lines that secrete NEO mAb were established with monoclonal antibody hybridoma tech2 nology. The immunological traits such as titer,affinity,sensitivity and specificity of the mAb were characterized. The re2 sults showed that NEO artificial antigen was synthesized successfully. Three hybridoma cell lines of 1E9、4E8 and 1G1 were screened for specificity to NEO. The indirect ELISA titer of the mAb were 1∶2. 56 ×103, 1 ∶1. 28 ×103 and 1∶5. 12 ×103 in supernatant,and 1∶5. 12 ×105 ∶2. 56 ×105 and 1∶1. 02 ×106 in ascites. The affinity constant (Ka) of 1G1 was about 3. 75 ×1010 (L/ mol). The mAb of 1G1 showed good sensitivity with an IC50 of 2. 57 ng/ mL to NEO. No cross2reactivity to other compounds was detected, snch as Gentamicin,Streptomycin,Oxytetracycline,Ciprofloxacin,Di2 floxacin,etc. The NEOmAb with high2titer,sensitivity and specificity had been generated,and could be used to establish the immunoassay of NEO residues in feed and animal food.

Key words: Neomycin, Monoclonal antibody, Immunological traits

CLC Number: 

• S859.8