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当前位置: 首页 > 文章 > 粮油产品T-2毒素单克隆抗体研制及应用 中国油料作物学报 2015,37 (5) 724-729
Position: Home > Articles > Preparation and application of a monoclonal antibody against T-2 toxin in grain and oil products Chinese Journal of Oil Crop Sciences 2015,37 (5) 724-729

粮油产品T-2毒素单克隆抗体研制及应用

作  者：
张亮;李培武;张奇;张文;杨青青;朱建国
单  位：
中国农业科学院油料作物研究所
关键词：
T-2毒素;单克隆抗体;间接竞争ELISA;检测
摘  要：
以T-2毒素与琥珀酸为原料,通过琥珀酸酐法合成出T-2毒素半抗原T-2HS,再通过活泼酯法,将T-2毒素半抗原分别与牛血清蛋白(BSA)和卵清白蛋白(OVA)共价偶联,合成出T-2毒素人工抗原T-2HSBSA和T-2HS-OVA,以T-2HS-BSA作为免疫原,免疫Balb/c小鼠,经细胞融合和阳性筛选,共获得4株T-2毒素单克隆抗体杂交瘤细胞株,分别是1D6、2A8、2C5和4G3。其中2C5分泌的抗体灵敏度(IC50)最高,达0.13μg/kg,特异性强,与HT-2的交叉反应率仅为4%,与FB1、DON、ZEN无交叉反应。进一步利用该抗体建立了间接竞争ELISA检测方法,最低检测限为0.015μg/kg,检测范围(IC20~IC80)为0.05~57.6μg/kg,该方法对大豆、玉米、花生等粮油产品的检测范围为0.5~576μg/kg,在线性范围内加标回收率为93.5%~107.5%。本研究建立的检测方法可用于大豆、玉米、花生等多种粮油产品中T-2毒素的检测。
译  名：
Preparation and application of a monoclonal antibody against T-2 toxin in grain and oil products
作  者：
ZHANG Liang;LI Pei-wu;ZHANG Qi;ZHANG Wen;YANG Qing-qing;ZHU Jian-guo;Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences;Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture;Key Laboratory of Detection for Mycotoxins,Ministry of Agriculture;Laboratory of Risk Assessment for Oilseeds Products ( Wuhan) ,Ministry of Agriculture;Quality Inspection and Test Center for Oilseeds Products,Ministry of Agriculture;
关键词：
T-2 toxin;;Monoclonal antibody;;Indirect competitive ELISA;;Detection
摘  要：
T- 2 toxin was reacted with succinic anhydride to prepare T- 2- hemisuccinate( T- 2HS). T-2HS was conjugated to bovine serum albumin( BSA) and ovalbumin( OVA) to obtain the conjugates T- 2HS-BSA and T- 2HS- OVA. As immunogen,T- 2HS- BSA was intradermally injected in Balb / c mice. Spleen cells and myeloma cells were fused and selected. 4 positive hybridoma cell lines were obtained,which were 1D6,2A8,2C5 and 4G3. The antibody from 2C5 had the best sensitivity with an IC50 of 0. 13μg·kg- 1. This antibody was against T- 2 particuarly,and the cross reactivity with HT- 2 was 4%. It had no cross reactivity with mycotoxins FB1,DON and ZEN. Indirect competitive ELISA was developed for T- 2 toxin detection,and the limit of detection was 0. 015μg·kg- 1. The detection range( IC20- IC80) of this method was 0. 05- 57. 6μg·kg- 1. The recoveries of spiked T- 2 toxin,in 0. 05- 57. 6μg·kg- 1,were 93. 5%- 107. 5%. It indicated that the above method could be used for T- 2 toxin detection in soybean,corn and peanut products.

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