食品科学 ›› 2009, Vol. 30 ›› Issue (9): 172-175.doi: 10.7506/spkx1002-6630-200909041

• 生物工程 • 上一篇    下一篇

赭曲霉毒素A单克隆抗体免疫亲和柱的制备

游淑珠1, 2, 3,许 杨1,2,*,邓舜洲1,2,黄志兵1,2   

  1. 1.南昌大学 食品科学与技术国家重点实验室 2. 南昌大学中德联合研究院 3. 珠海出入境检验检疫局技术中心
  • 收稿日期:2008-08-09 修回日期:2009-01-08 出版日期:2009-05-01 发布日期:2010-12-29
  • 通讯作者: 许杨 E-mail:xuyang@nuc.edu.cn,xuyang1951@163.com
  • 基金资助:

    教育部“长江学者和创新团队发展计划”项目(IRT0540)

Development of Immunoaffinity Column of Anti-ochratoxin A Monoclonal Antibody

YOU Shu-zhu1, 2, 3 XU Yang1,2,* DENG Shun-zhou1,2 HUANG Zhi-bing1,2   

  1. 1.State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China
    2.Sino-Germany Joint Research Institute, Nanchang University, Nanchang 330047, China
    3.Technical Center of Zhuhai Entry-Exit Inspections and Quarantine Bureau, Zhuhai 519015, China
  • Received:2008-08-09 Revised:2009-01-08 Online:2009-05-01 Published:2010-12-29
  • Contact: XU Yang E-mail:xuyang@nuc.edu.cn,xuyang1951@163.com

摘要:

制备了赭曲霉毒素A(OTA)单克隆抗体免疫亲和柱,并用间接竞争ELISA和HPLC法评价了免疫亲和柱的性能,其柱容量(结合OTA的能力)约为200ng,加标回收率为90.38%~100.1%,可反复使用3次。 建立了免疫亲和柱-HPLC联用分析谷物中OTA的方法,最低检出限为0.2μg/kg,线性范围为0.6~400μg/kg,OTA加标量为1~10μg/kg时谷物样品中的回收率为78.7%~87.1%,变异系数小于6.5%。用此法检测了大米、小麦、玉米和玉米饲料等15份市售样品,检出率为46.7%,其中OTA的最高含量为0.785μg/kg。

关键词: 赭曲霉毒素A, 单克隆抗体, 免疫亲和柱

Abstract:

An immunoaffinity column (IAC) was prepared with anti-ochratoxin A (OTA) monoclonal antibody  and the efficiency of IAC was analyzed by indirect competitive enzyme-linked immunosobent assay (IC-ELISA) and high performance liquid chromatography (HPLC). Results indicated that this IAC had a high column capacity of binding OTA (200 ng) and the recovery ranging from 90.38 % to 100.1 %, and could be used repeatedly 3 times. IAC linked with HPLC was used to detect OTA contents in 15 cereal samples. The limit of detection (LOD) of OTA by this method was 0.2  μg/kg. Samples with OTA levels from 0.6 to 400  μg/kg could be quantitified by this method. The recovery of spiked OTA at levels of 1 to 10 μg/kg ranged from 78.7 % to 87.1 % with coefficients of variation (CVs) less than 6.5 %. This method was applied to detect 15 saled cereal samples. The positive detection rate was 46.7 %, and the highest OTA content detected was 0.785 μg/kg.

Key words: ochratoxin A, monoclonal antibody, immunoaffinity column

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