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Preparation of synthetic antigen and monoclonal antibody for indirect competitive ELISA of citrinin

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Citrinin (CIT)-protein synthetic antigen was prepared by conjugating CIT with keyhole limpet hemocyanin by 1,4-butanediol diglycidyl ether. The ultraviolet and infrared spectrometry analyses indicated that CIT was conjugated with the carrier protein successfully. By fusion and cloning, a hybridoma cell line K2-F3 which stably secreted the highly specific monoclonal antibody (McAb) against CIT was obtained. The indirect competitive ELISA showed that the IC50 value was 0.3 ng/mL and the limit of detection, 0.01 ng/mL. Cross-reactivity was less than 0.01% with four mycotoxins, aflatoxin B1, ochratoxin A, zearalenone, deoxynilvalenol and three pigments, rubropunctatine, rubropunctamine, monascin. The mean recovery of citrinin from artificially contaminated red yeast rice was from 89 to 92%, with CVs from 6.8 to 9.0%. The ELISA developed in this study can accurately determine CIT in artificially contaminated red yeast rice.
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Keywords: ELISA; citrinin; monoclonal antibody; synthetic antigen

Document Type: Research Article

Affiliations: 1: College of Chemistry & Chemical Engineering,Fuzhou University, Fuzhou350002, P.R. China 2: Fujian Key Laboratory of Medical Devices and Pharmaceutical Technology,Fuzhou University, Fuzhou,350002, P.R. China

Publication date: June 1, 2012

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