Preparation of monoclonal antibodies against a derivative of semicarbazide as a metabolic target of nitrofurazone

摘要

Monoclonal antibodies(McAb)were produced to detect semicarbazide(SEM),a metabolite as a marker residue of nitrofurazone in animal food production.A carboxyphenyl derivative(CPSEM)of SEM was synthesized following derivatisation with 4-carboxybenzaldehyde(CBA).CPSEM was purified by recrystallization and conjugated to bovine serum albumin(BSA)or ovalbumin(OVA)as immunogen or coating antigen,respectively.Hybridomas were obtained by fusing mouse myeloma cells SP2/0 with splenocytes from the mice immunized with CPSEM-BSA.Hybridomas 1F10 and 4F2 secreting antibodies against CPSEM were obtained and subcloned.Ascites of monoclonal antibodies were prepared by injecting 1 x 10~6 cells of hybridoma 1F10 into mice abdomen.McAb obtained from hybridoma 1F10 was highly specific for CPSEM and had no cross-reaction with various nitrofuran metabolites and a range of veterinary drugs.The sensitivity of the McAb to SEM was 0.01 ng mL~(-1)and the IC_(50)value was 1.3 ng mL~(-1)(SEM in the form of CPSEM).McAb 1F10 is suitable to develop an immunoassay for SEM with sufficient sensitivity for monitoring nitrofurazone residues.