核农学报2013，Vol.27Issue(1)：88-92,5.
庆大霉素单克隆抗体的研制及ELISA分析方法的建立
PREPARATION OF MONOCLONAL ANTIBODY AND DEVELOPMENT OF AN INDIRECT COMPETITIVE ELISA METHOD FOR GENTAMICIN
摘要
Abstract
One hybridoma cell line (6H8) secreting monoclonal antibody (McAb) against gentamicin was produced by fusing mouse myeloma cells ( SP2/0) with spleen cells from BALB/C which immunized by the artificial gentamicin antigen conjugated with bovine serum albumin ( BSA). Isotype and subclass of the monoclonal antibody secreted from the hybridoma cell line (6H8) was classified as IgGl. The light chain of the McAb was identified to be Κ chain. The McAb obtained could specifically react with gentamicin and its titre of ascitic fluid detected by indirect ELISA was up to 1 × 10-7. The result of specificity analysis indicated that the McAb had no cross-reactivity with analogues of gentamicin. Based on the producted McAb, an indirect competitive ELISA was established, and its linear regression equation was y = 16. 1221n (x) -2. 0143 (R2 =0. 9934). Inhibition rate analysis showed that IC50 and IC20 values were 25. 2 μg·L-1 and 3. 9 μg·L-1 gentamicin in PBS buffer, respectively. The mean recovery of gentamicin spiked in milk was from 91% to 110%. The producted anti-gentamicin McAb and established competitive ELISA method could lay the foundation for rapid detection of gentamicin residue.关键词
庆大霉素/单克隆抗体/竞争ELISAKey words
Gentamicin/ monoclonal antibody/ competitive ELISA引用本文复制引用
金仁耀,吴建祥..庆大霉素单克隆抗体的研制及ELISA分析方法的建立[J].核农学报,2013,27(1):88-92,5.基金项目
浙江省科技计划项目(2011C22078) （2011C22078）
