铬离子单克隆抗体的制备及食品总铬含量检测胶体金免疫层析试纸条的研制  

Preparation of monoclonal antibody against trivalent chromic ion and development of colloidal gold immunochromatographic assay for detecting total chromium content in food

作　　者：王亚楠[1] 王晓斐[2] 丁菡[1] 张海棠[1] 范国英[1] 王自良[1] 

机构地区：[1]河南科技学院动物科学学院,河南新乡453003 [2]河南科技学院新科学院,河南新乡453003

出　　处：《食品工业科技》2016年第18期63-69,共7页Science and Technology of Food Industry

基　　金："十二五"国家科技支撑计划项目(2011BAK10B01);"十二五"国家科技支撑计划项目(2014BAD13B05)

摘　　要：目的:制备三价铬离子螯合物单克隆抗体(Cr^(3+)-EDTA m Ab),研制胶体金免疫层析试纸条(Cr-Strip)。方法:用异硫氰酯法合成Cr^(3+)-i EDTA-BSA,UV和ICP-AES进行鉴定;细胞融合技术筛选Cr^(3+)-EDTA m Ab细胞株,体内诱生腹水法制备Cr^(3+)-EDTA m Ab,并对其特性进行分析;应用Cr^(3+)-EDTA m Ab研制Cr-Strip,并测定其性能。结果:筛选出3株杂交瘤,最好的是2A11G5,Ka为2.69×109L/mol,对Cr^(3+)-EDTA的IC50为9.84μg/L,与其他重金属离子无交叉反应;Cr-Strip的检测时间为10 min,检测限为5μg/L,其检测结果与ICP-AES符合率为100%。结论:制备出了高质量的Cr^(3+)-EDTA m Ab,研制出更为灵敏的Cr-Strip。Object： To prepare monoclonal antibody against Cr3＋ chelate EDTA complex （ Cr3＋ - EDTA mAb） and develop colloidal gold immunochromatographic assay for detecting total chromium content in food （Cr-Strip）. Method：The isothiocyanate method was employed to synthesize Cr3＋ iEDTA-BSA, which was identified by UV and ICP-AES.The hybridoma lines that secrete Cr3＋-EDTA mAb were established by cell fusion and Cr3＋ -EDTA mAb were induced from in vivo method and their immunological properties were analyzed.Based on Cr3＋- EDTA mAb, Cr-Strip was developed and its traits were verified.Results：Three hybridoma lines were screened out and the best one was 2A11G5.The Ka of Cr3＋- EDTA mAb was 2.69 × 109 L/mol and its IC50 against Cr3＋- EDTA was 9.84 μg/L and it had little or no cross-reactivity with other metal ion. The Cr-Strip could be accomplished qualitative detection of Cr3＋- EDTA in 10 minutes,and its detection limit was 5 μg/L,its coincidence rate was 100% compared with ICP-AES.Conclusion：The high quality Cr3＋ -EDTA mAb had been generated and the Cr-Strip which was more sensitive had been developed successfully.

关 键 词：Cr^(3+) 免疫原 单克隆抗体 胶体金免疫层析 总铬含量 

分 类 号：TS207.3[轻工技术与工程—食品科学]