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  • [Preparation of anti-zearalenone monoclonal antibody and preliminary establishment of colloidal gold immunochromatographic assay for zearalenone].

[Preparation of anti-zearalenone monoclonal antibody and preliminary establishment of colloidal gold immunochromatographic assay for zearalenone].

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (2013-07-11)
Miner Luo, Yong Tang, Junjian Xiang, Xiaoli Zhang, Qiangqiang Fu, Hong Wang
초록

To prepare the monoclonal antibody against zearalenone (ZEN) toxin and preliminarily establish the colloidal gold immunochromatographic detection method for ZEN. The artificial antigen ZEN-BSA and ZEN-OVA were prepared by active ester method. Mice were immunized with ZEN-OVA and monoclonal antibodies against ZEN were prepared by regular cell fusion and subcloning approach. The titer, subtype and specificity of the antibodies were identified by ELISA. To establish a method of colloidal gold immunochromatographic assay (GICA) for the determination of ZEN, colloidal gold binding cushion was coated with anti-ZEN monoclonal antibody-colloidal gold complex, and the synthetic ZEN-BSA and goat anti-mouse immunoglobulins were sprayed on cellulose nitrate film to form the test (T) band and control (C) band. Identified by SDS-PAGE and UV spectroscopy, the artificial antigen ZEN-BSA and ZEN-OVA were successfully coupled respectively. One hybridoma line (1G4) which could secrete monoclonal antibody specifically against ZEN was obtained by three-time cloning. The ascites titer of this monoclonal antibody reached 1:1.6×10(5);. The antibody subtype was IgG2b. The IC50; to ZEN was 10.2 ng/mL, and the detction limit was 0.58 ng/mL. In addition, the mAb was proved highly specific for ZEN. ZEN metabolites α-zearalanol, β-zearalanol, zearalanone and other similar toxins deoxynivalenol, fumonisin B1, ochratoxin A showed very low or no cross-activity with the monoclonal antibody. The whole assay using the prepared colloidal gold immunochromatographic strip could be finished in 5 min, which could be read by naked eyes. The limit of detection was 100 ng/mL. The anti-ZEN mAb was successfully prepared. And the gold immunochromatographic assay(GICA) for ZEN was preliminarily established with the limit of detection being 100 ng/mL.

MATERIALS
제품 번호
브랜드
제품 설명

Supelco
Zearalenone solution, 100 μg/mL in acetonitrile, analytical standard
Sigma-Aldrich
Zearalenone, fungal mycotoxin