Preparation of monoclonal antibodies against Carprofen
10.3969/j.issn.1000-484X.2017.11.015
- VernacularTitle:卡洛芬单克隆抗体的制备
- Author:
Hua Yan XIONG
1
;
Zhong Ai GAO
;
Yi WANG
Author Information
1. 湖北省中西医结合医院
- Keywords:
Carprofen;
Monoclonal antibody;
ELISA;
Colloidal gold
- From:
Chinese Journal of Immunology
2017;33(11):1673-1677
- CountryChina
- Language:Chinese
-
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.
