分析化学2016，Vol.44Issue(9)：1385-1393,9.DOI:10.11895/j.issn.0253-3820.160360
孔雀石绿单克隆抗体制备和酶联免疫检测方法的建立
Synthesis of Novel Hapten and Development of Monoclonal Antibody-based Enzyme-Linked Immunosorbent Assay for Malachite Green in Fish Samples
摘要
Abstract
To produce specific antibodies against malachite green ( MG) , one special hapten was synthesized and characterized, and conjugated to carrier protein as immunogen. The immunogen showed excellent reactogenicity and immunogenicity. One specific monoclonal antibody (mAb, named MG-DA4-C7) with high sensitivity and specificity for MG in indirect competitive enzyme-linked immunoassay ( icELISA ) was screened. The isotype was IgG1 and the light chain was κ type. After optimization of ELISA conditions, the proposed icELISA showed a 50% inhibition value ( IC50 ) of 0. 96 μg/L, a linear range ( IC20-IC80 ) of 0. 1-8. 1 μg/L and a limit of detection ( LOD, IC10 ) of 0. 05 μg/L for determination of MG. The assay showed cross-reactivity of 18. 1%, 26. 5% with crystal violet and brilliant green, respectively, and negligible cross-reactivity with other metabolites of MG (<0 . 1%) . The average recoveries of MG from spiked fish samples were from 87. 3% to 107. 3%. Good correlation (R2=0. 999) was obtained between the results of icELISA and those of liquid chromatography-tandem mass spectrometry analysis. The proposed icELISA is suitable for the determination of MG in fish samples in a simple and sensitive manner.关键词
孔雀石绿/半抗原/单克隆抗体/抗原识别/酶联免疫分析Key words
Malachite green/Hapten/Monoclonal antibody/Antigenic-determinant/Enzyme-linked immunoassay引用本文复制引用
王宇,杨金易,徐振林,戚平,沈玉栋..孔雀石绿单克隆抗体制备和酶联免疫检测方法的建立[J].分析化学,2016,44(9):1385-1393,9.基金项目
本文系国家自然科学基金(No.31371769),广东省科技计划项目(Nos.2014A020219008,2013B040402006,2016A040403077),广东省自然科学基金项目(Nos.2013030013338,2014A030306026)项目资助。 （No.31371769）
