A sensitive immunoaffinity column-linked indirect competitive ELISA for ochratoxin A in cereal and oil products based on a new monoclonal antibody.

摘要

A new sensitive monoclonal antibody (mAb) 1H2 against ochratoxin A (OTA) was reported herein. This mAb belonged to the immunoglobulin G1 (k chain) isotype. In the optimized indirect competitive enzyme-linked immunosorbent assay (icELISA), 1H2 showed a 50 % inhibition concentration (IC₅₀) value of 0.058 ng/mL and a detection limit (IC₁₀) of 0.001 ng/mL. The cross-reactivity of 1H2 with ochratoxin B, aflatoxins, deoxynivalenol, zearalenone, T-2 toxin, or fumonisins was below 0.3 %. Based on this mAb, an immunoaffinity column (IAC)-linked icELISA was developed for OTA detection in the cereal and oil products. The working range of the assay for solid sample was 0.36-16m mug/kg. The recoveries from spiked samples of IAC-linked icELISA ranged from 83 to 101 %. These recoveries were much higher than those of icELISA (21-78 %) and in good agreement with those obtained by using the standard high-performance liquid chromatography method (87-110 %). The results indicated that the mAb 1H2 had the values for studies of OTA in the crude agricultural products. copyright Springer Science+Business Media New York 2013.