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

王亚楠; 王晓斐; 丁菡; 张海棠; 范国英; 王自良

doi:10.13386/j.issn1002-0306.2016.18.004

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

摘要

摘要: 目的:制备三价铬离子螯合物单克隆抗体(Cr³⁺-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Cr³⁺-i EDTA-BSA,UV和ICP-AES进行鉴定;细胞融合技术筛选Cr³⁺-EDTA m Ab细胞株,体内诱生腹水法制备Cr³⁺-EDTA m Ab,并对其特性进行分析;应用Cr³⁺-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Cr³⁺-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Cr³⁺-EDTA m Ab,研制出更为灵敏的Cr-Strip。

Abstract: Object: To prepare monoclonal antibody against Cr³⁺chelate EDTA complex( Cr³⁺- EDTA m Ab) and develop colloidal gold immunochromatographic assay for detecting total chromium content in food( Cr- Strip).Method: The isothiocyanate method was employed to synthesize Cr³⁺- i EDTA- BSA,which was identified by UV and ICP- AES.The hybridoma lines that secrete Cr³⁺- EDTA m Ab were established by cell fusion and Cr³⁺- EDTA m Ab were induced from in vivo method and their immunological properties were analyzed. Based on Cr³⁺- EDTA m Ab,Cr- Strip was developed and its traits were verified.Results: Three hybridoma lines were screened out and the best one was 2A11G5. The Kaof Cr³⁺- EDTA m Ab was 2.69 × 109 L / mol and its IC50 against Cr³⁺- EDTA was9.84 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 m Ab had been generated and the Cr- Strip which was more sensitive had been developed successfully.