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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. [ABSTRACT FROM PUBLISHER]

ABSTRACT Citrinin (CIT)-protein artificial antigen was prepared by conjugating CIT with keyhole limpet hemocyanin by 1,4-butanediol diglycidyl ether. By immunization and fusion, a hybridoma cell line named K2-F3, which stably secreted the monoclonal antibody (McAb) against CIT was obtained. The titer of the CIT- specific McAbs purified by affinity chromatography came to 1:1.38 × 105. The indirect competitive enzyme-linked immunosorbent assay (ELISA) showed that the detection limit of CIT was 0.01 ng/mL, with a good linearity ranging from 0.01 to 1.0 ng/mL. The McAb was highly specific and cross-reactivity rate was less than 0.01%. The data showed that the ELISA developed could accurately determine CIT in the real contaminated red yeast rice samples. The systematic error was low with the coefficient of variation from 2.91 to 8.53% by the ELISA. PRACTICAL APPLICATIONS CIT is a toxic fungal metabolite. It often occurs in naturally contaminated commodities, such as in corn, barley, wheat, apple and some foods . CIT is hepatonephrotoxic and implicated in disease outbreaks in animals and humans. Nowadays, food security problem due to the contamination of CIT has been a growing concern for the government and the public, especially in Asian countries. In this work, a high-specificity McAb against citrinin was obtained. The indirect competitive ELISA based on the obtained McAb exhibited a high sensitivity and precision. The detection limit of CIT was 0.01 ng/mL. The indirect competitive ELISA developed could determine CIT with a satisfactory precision for the real red yeast rice samples. This work would be helpful for establishing the technology and developing the kit to determine CIT-contaminated samples by ELISA. [ABSTRACT FROM AUTHOR]