中国食品学报2013，Vol.13Issue(1)：145-152,8.
玉米赤霉烯酮单克隆抗体制备及免疫分析
Preparation of Monoclonal Antibody and Detection of Zearalenone by Enzyme-linked Immunosorbent Assay
摘要
Abstract
Zearalenone (ZEN) was coupled with bovin serum albumin(BSA) and ovalbumin(OVA) to prepare im-munogen ZEN—BSA and coating antigen ZEN-OVA by ester activation method. The two antigens were identified by UV Scan and SDS-PAGE. The spleen cells of BALB/c mice immunized by ZEN-BSA were fused with SP2/O myeloma cells, and hybridomas secreting antibodies against ZEN were selected and cloned. A stable hybridoma cell line 3D8 that secret MAb of subclasses IgGl, kappa light chain were established. The titer of the MAb determined by indirect ELISA was 1:2.048×106 in ascites. Based on the ascites MAb, an indirect competitive ELISA (ic-ELISA) method was developed for the quantitative detection of ZEN. The ic-ELISA had a good sensitivity with an IC50 of 22.89 pg/mL, a detection limit of 10.07 pg/mL, and 95%, 8%, 12%, 6%, 5% cross-reactivitives to α-zearalenol, β-zearalenol, zearalanone, α-zear-alanol, β-zearalanol respectively, and scarcely showed cross-reactivity to others. The spiked recovery of this method ranged from 86.4% to 104.8% in maize, barley, wheat and oat samples. It is indicated that the developed method in this paper was sensitive and convenient in preliminary screening of ZEN.关键词
玉米赤霉烯酮/单克隆抗体/间接竞争ELISAKey words
Zearalenone/ monoclonal antibody/ indirect competitive ELISA引用本文复制引用
李沐洁,张明洲,奚茜,陈宗伦,王唯芬,龚云飞..玉米赤霉烯酮单克隆抗体制备及免疫分析[J].中国食品学报,2013,13(1):145-152,8.基金项目
浙江省重大科技专项(2006C12102) （2006C12102）
杭州市重大科技创新专项(20083212A25) （20083212A25）
浙江省重点科技创新团队(2010R50028) （2010R50028）
浙江省科技成果转化项目(2011E61018) （2011E61018）
浙江省重大科技专项重点项目(2012C12013-3) （2012C12013-3）
