玉米赤霉烯酮单克隆抗体酶联免疫测定方法的建立及初步应用

ESTABLISHMENT AND PRELIMINARY APPLICATION OF McAb-CIEIA FOR DETERMINATION OF ZEARALENONE

用ZEN-BSA人工抗原免疫BALB/c鼠,经融合、筛选和克隆化得到可稳定分泌抗ZEN单克隆抗体的杂交瘤细胞株ZEN-1C6。ZEN-1C6属IgG_1,纯化腹水抗体效价为10^(-5),与5种衍生物的交叉反应系数为0.16～1.20％。用ZEN-1C6建立了检测食品(玉米、小麦、大米)中玉米赤霉烯酮的CIEIA法。该法检测纯毒素的线性范围为5～1000ng/ml,最低检出浓度为0.1ng/ml,平均回收率为84.0～105.5％。用该法测定了81份样品,均有ZEN毒素检出。

In the present experiment, a hybridoma cell line secreting monoclonal antibody against zearalenone (ZEN) was established by fusion of Sp-2 /0-Ag-14 and spleen cells from BALB /c mice immunized with ZEN-BSA conjugate. The McAb, designated ZEN-1C6, was of the subtype IgG1 and the ascitic fluid of which was 10-5 by indirect ELISA. The cross - reactivities of this McAb for zearalanone, α-zearalenol, β - zearalenol, α-zearalanol, and β - zearalanol were 1.20, 1.14, 0.16, 1.03, and 0.40%, respectively, of that found for ZEN. An competitive inhibition enzyme immunoassay (CIEIA) was developed for the detection of ZEN in food (including corn, wheat and rice) using ZEN-1C6. The linear range for ZEN by the method was between 5 and 1000 ng /ml and the minimum detectable concentration was 0. 1ng /ml. The mean recovery rates from sample which artificially contaminated with 10 -1000μg /kg of ZEN were 84.0 - 105.5%, A total of 81 samples collected from market were analyzed with the CIEIA, and the concentration of ZEN ranged from 0.2 to 125.8μg /kg.