Preparation of immunoaffinity column of zearalenone
Zhang, Y.; Huang, Z-bing.; Deng, S-zhou.; Xu, Y.
Zhonghua Yu Fang Yi Xue Za Zhi 41(2): 110-113
2007
ISSN/ISBN: 0253-9624 PMID: 17605236 Document Number: 611270
Objective To prepare immunoaffinity column of zearalenone. Methods The zearalenone immunoaffinity column(IAC) was prepared by coupling CNBr-activated Sepharose 4 Fast Flow (4FF) with the anti -zearalenone monoclonal antibody which was purified by caprylic acid -ammonium sulfate method. The coupling reaction was identified by UV-absorbance measurements, and the IAC prepared was evaluated by indirect-competition ELISA and HPLC. Results The column capacity was determined to be 0. 40 mu g when using 0. 5 ml of CNBr activated Sepharose 4FF and 350 eta g of purified anti-zearalenone monoclonal antibody. The mean true recoveries were in the range 76. 33 % - 90. 10 % and RSD was 6. 68 % - 10. 93 % at levels of 60 mu g/kg - 300 mu g/kg. 30 samples of wheat and maize were detected by the anti-ZEN IAC produced by the laboratory, 17 samples were observed to be contaminated in a comparable range from 31. 33 mu g/kg - 377. 84 mu g/kg. Detection limit based on a signal-to-noise ratio 3: 1 was 10. 00 mu g/kg for ZEN in wheat and maize. Conclusion IAC, a simple separating method which is used in ZEN extraction from cereals, is able to purify and condense ZEN in one step. The cost of detection can be lowered down because the IAC developed is hopefully to substitute the imported IAC.