食品科学 ›› 2009, Vol. 30 ›› Issue (12): 258-261.doi: 10.7506/spkx1002-6630-200912059

• 分析检测 • 上一篇    下一篇

柠檬黄色素免疫学检测方法的研究

任立松1 , 2,卢士英1,周 玉1,郭德军1,李岩松1,孟宪梅1,柳增善*   

  1. 1.吉林大学人兽共患病研究所 2.新疆军区疾病预防控制中心
  • 收稿日期:2008-05-06 修回日期:2008-12-22 出版日期:2009-06-15 发布日期:2010-12-29
  • 通讯作者: 柳增善* E-mail:zsliu1959@sina.com

Immunoassay Determination of Tartrazine

REN Li-song1,2,LU Shi-ying1,ZHOU Yu1,Guo De-jun1,LI Yan-song1,MENG Xian-mei1,LIU Zeng-shan*   

  1. (1. Institute of Zoonosis, Jilin University, Changchun 130062, China;
    2. Xinjiang Military Center for Disease Control and Prevention, Urumqi 830011, China)
  • Received:2008-05-06 Revised:2008-12-22 Online:2009-06-15 Published:2010-12-29
  • Contact: LIU Zeng-shan* E-mail:zsliu1959@sina.com

摘要:

将柠檬黄分子与卵清蛋白(OVA)、牛血清白蛋白(BSA)偶联为检测原和免疫原,免疫Balb/c 小鼠,通过杂交瘤技术获得分泌抗柠檬黄单克隆抗体杂交瘤细胞株3D4,纯化腹水得到单克隆抗体,建立间接竞争ELISA 检测方法,回归方程和相关系数分别为:y= - 40.134x+133.46,R2=0.9692,最低检出浓度为0.36μg/L。该方法与柠檬黄色素常用的检测方法薄层分析及分光光度计法相比,检测限低、耗时短、成本低,适用于大批量样品检测。

关键词: 柠檬黄色素, 单克隆抗体, 酶联免疫吸附反应, 检测

Abstract:

Tartrazine was coupled separately with bovine serum albumin (BSA) and ovalbumin (OVA) by 1-ethyl-3-(3- dimethylaminopropyl) carbodiimide (EDC) to prepare complete antigens. Then hybridoma cell lines secreting monoclonal antibody (McAb) against tartarzine were obtained by immunization of BALB/C mice with prepared BSA- tartarzine and then fusion between SP2/O myeloma cells and splenocytes from the immunized mice, and the McAb against tartarzine was prepared. Finally, an indirect competitive ELISA method was established for determination of tartrazine. The regression equation was y =-40.134x + 133.46, with a determination coefficient of 0.9692. The limit of detection was 0.36 μg /L. Compared to the widely used methods such as TLC and spectrophotometric assay, this new method is found to be time-saving, easy to perform, highly specific and sensitive. It was suitable for the determination of tartarzine-containing samples in large quantity.

Key words: tartarzine, McAb, ELISA, detection

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