ABSTRACT
Objective:Preparation of monoclonal antibodies against Carprofen,which lays a foundation for the rapid detection of Carprofen.Methods:Via an active ester method,Carprofen was conjugated to bovine serum albumin (BSA) as immune antigen and ovalbumin (OVA) as detection antigen,respectively.Hybridomas were obtained by fusing mouse myeloma cells SP2/0 with splenocytes from the mice immunized with Carprofen-BSA.Hybridomas 51C3 secreting antibodies against Carprofen were obtained and subcloned.Results: Ascites of monoclonal antibodies (McAb) were prepared by injecting cells of hybridoma 51C3 into mice abdomen.The titer of purified McAb was 3.2×105and the McAb was IgG1 subtype.McAb was highly specific for Carprofen and the cross-reactivity (CR50%) was less than 0.04% with Ibuprofen, Ketoprofen, Naproxen, Feinuoluofen, Indoprofen, Fenbufen respectively.The sensitivity of the McAb to carprofen was 0.32 ng/ml and the IC50value was 0.882 ng/ml.The recoveries of Carprofen in the pork samples were from 81.4% to 104.4 % and coefficients of variation were from 16.3% to 25.8%.Conclusion:All results indicate that the McAb 51C3 is suitable to develop an immunoassay colloidal gold rapid dipstick test.
