苏丹红Ⅰ、Ⅲ和对位红竞争ELISA检测方法的初步建立

食品科学 ›› 2008, Vol. 29 ›› Issue (8): 523-525.

苏丹红Ⅰ、Ⅲ和对位红竞争ELISA检测方法的初步建立

唐勇, 琚春梅, 魏钟杰,向军俭

暨南大学抗体工程研究中心；华南农业大学动物医学院；广州市江森生物科技有限公司

出版日期:2008-08-15 发布日期:2011-08-26

Primary Development of Competitive ELISA for Sudan Ⅰ,Ⅲ and Para Red Detection

TANG Yong, JU Chun-Mei, WEI Zhong-Jie, XIANG Jun-Jian

1.Center of Antibody Engineering,Jinan University,Guangzhou 510632,China;2.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;3.Guangzhou Jiangsen Biotech Limited Company,Guangzhou 510632,China

Online:2008-08-15 Published:2011-08-26

摘要/Abstract

摘要： 以4-氨基苯甲酸重氮化后与2-萘酚反应,合成苏丹红Ⅰ号的衍生物(CSD Ⅰ),以活性脂法将CSD Ⅰ与蛋白载体偶联,制备CSD Ⅰ-BSA免疫抗原与CSD Ⅰ-OVA检测抗原。以CSD Ⅰ-BSA免疫Balb/c小鼠,适当免疫后取脾细胞与SP2/0融合,筛得抗CSD Ⅰ的单克隆抗体细胞株5H3。以该细胞株生产抗体与CSD Ⅰ-OVA建立CSD Ⅰ的间接竞争ELISA检测方法,该方法对苏丹红Ⅰ、Ⅲ和对位红的灵敏度约为0.1ng/ml,IC50分别为1.03、1.13、0.89ng/ml,与其他染料交叉反应率低。该ELISA检测方法灵敏、特异,可用于苏丹红Ⅰ、Ⅲ和对位红的快速检测。

关键词: 苏丹红, 抗体, 检测方法

Abstract: A derivative(CSD Ⅰ) of Sdan I was synthesized following the diazonium coupling reaction of 2-naphthol with 4-carboxybenzenediazonium ion obtained from 4-aminobenzoic acid. CSD Ⅰ was conjugated to bovine serum albumin(BSA) or ovalbumin(OVA) as immunogen or coating antigen,respectively. Hybridomas were obtained by fusing mice myeloma cells SP2/0 with splenocytes from the mice immunized with CSD Ⅰ-BSA. Hybridoma 5H3 secreting antibodies against CSD Ⅰ was obtained. The competitive ELISA was developed based on CSD Ⅰ-OVA and the antibodies against CSD Ⅰ. The sensitivity of ELISA to sudan Ⅰ,Ⅲ and para red was approximately 0.1 ng/ml,and their IC50 values are 1.03,1.13,0.89 ng/ml,respectively. The ELISA is high specific for Sudan Ⅰ,Ⅲ and para red and low cross reaction with other Sudan red and related dyes. The ELISA method is sensitive and specific which can be used for fast screening of illegal Sudan Ⅰ,Ⅲ and para red.

Key words: Sudan red, antibody, detection method

唐勇, 琚春梅, 魏钟杰, 向军俭. 苏丹红Ⅰ、Ⅲ和对位红竞争ELISA检测方法的初步建立[J]. 食品科学, 2008, 29(8): 523-525.

TANG Yong, JU Chun-Mei, WEI Zhong-Jie, XIANG Jun-Jian. Primary Development of Competitive ELISA for Sudan Ⅰ,Ⅲ and Para Red Detection[J]. FOOD SCIENCE, 2008, 29(8): 523-525.

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