摘要
采用优化的碳二亚胺法制备诺氟沙星(NOR)人工抗原,紫外扫描和红外光谱图表明人工抗原偶联成功。采用细胞融合技术筛选抗NOR单克隆抗体(NORmAb)杂交瘤细胞株,体内诱生腹水法制备NORmAb,建立间接竞争ELISA(icELISA)标准曲线。结果表明,筛选的3株杂交瘤细胞分泌的抗体为IgG1亚型,具k轻链;建立的icELISA标准曲线在PBS中的的线性检测范围为0.04~43.6ng/mL,IC50值为0.62ng/mL,检测限(LOD)为0.02ng/mL;抗体与环丙沙星(87.2%)、培氟沙星(76.9%)、依诺沙星(66.8%)、恩诺沙星(48.6%)和洛美沙星(36.6%)有较高的交叉反应率;禽肉基质中NOR添加回收率满足检测要求。
Modified EDC method was employed to synthesize the artificial antigen of norfloxacin(NOR),which was proved by Ultraviolet absorbance and IR spectra.Cell fusion technology was used to screen anti-NOR hybridoma lines,and monoclonal antibodies(mAbs)were produced by using ascitic fluid in mice.Based on this,an indirect competitive ELISA standard curve was established.This assay was sensitive and had a linear range from 0.04~43.6ng/mL(R2=0.9655),with IC50 and LOD values of 0.62ng/mL and 0.02ng/mL,respectively.The produced mAbs were of the IgG1 isotype with a k light chain,and mAb from N7B2 exhibited high cross-reactivities to ciprofloxacin(87.2%),pefloxacin(76.9%),enoxacin(66.8%),enrofloxacin(48.6%)and lomefloxacin(36.6%).The spiking tests indicated that the established icELISA method could be used for determination of NOR residue in poultry.
出处
《食品工业科技》
CAS
CSCD
北大核心
2011年第9期210-213,共4页
Science and Technology of Food Industry
基金
十一五国家科技支撑计划重大项目(2006BAK02A21/1)
河南省教育厅科技攻关项目(2011A230003)