Preparation Of Monoclonal Antibody To Sulfachlorpyrazine Specialty

Sulfonamides(SAs) is a kind of antibiotics with broad antibacterial spectrum andwide application. Irrational use of sulfa drugs in food production animals will lead todrug residues in animal tissue, which will ultimately do harm to people’s health. Inmany countries, the maximum residue level (MRL) of sulfonamides in food waslimited, and in some countries even regulate that sulfonamides can not be detected infood. Immunoassay technology, a veterinary drug residue detection technology withhigh sensitivity and with fast, convenient features, is especially suitable for sitescreening and large sample size analysis. Monoclonal antibody is the core componentof immunoassay technology. The purpose of this study is to produce monoclonalantibodies specific for SPZ artificial antigen using monoclonal antibody technology,and to provide a basis for rapid immunological test.In this study, BSA and OVA were used as carrier protein. With diazotizationmethod, the artificial antigen of SPZ was prepared successfully. Using UVspectrophotometry, SDS-PAGE and immunological detection, the results showed:artificial antigen absorption peaks are offset or superimposedand; the migration speedof the carrier protein is greater than the complete antigen; the serum titer of No.1andNo.2mouse can reach1:4000. These results proved the preparation of artificialantigens is successful.Using synthesized complete antigens BSA-SPZ immunized the BALB/c mice,screened mice with high titer serum titer, using hybridoma technology culturedimmune spleen cells with myeloma cells and got a total of128strains of hybridomacell lines. The hybridoma cell lines were preliminary screened by indirect ELISA andabtained3-positive cell lines, the rates of cell fusion were63.54%and69.79%, thepositive rates were1.5%and2.8%respectively. After three times clone, cell linessecreting anti SPZ monoclonal antibody was obtained, the antibody have a goodsensitivity and strong specificity.