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对位红抗原合成和多克隆抗体的制备
引用本文:程玛丽.对位红抗原合成和多克隆抗体的制备[J].中国农学通报,2011,27(14):82-86.
作者姓名:程玛丽
作者单位:1. 北京农学院动物科学技术学院,北京102206;中国农业科学院农业质量标准与检测技术研究所,北京100081
2. 中国农业科学院农业质量标准与检测技术研究所,北京,100081
3. 北京农学院动物科学技术学院,北京,102206
基金项目:北京市科学技术委员会项目
摘    要:制备了一种特异性强的抗对位红的多克隆抗体。采用混合酸酐法将活化的对位红半抗原与阳离子化牛血清白蛋白(cBSA)偶联成免疫原,免疫大白兔制备多克隆抗体,利用紫外分光光度计分析结合物的结合情况,间接ELISA和间接竞争ELISA分析抗体特性。紫外扫描分析的免疫原对位红与蛋白结合的偶联率为14:1,间接竞争ELISA法分析多克隆抗体效价达到1×106,50%抑制率检测限为7.43 ng/mL,检测的对位红线性范围在0.1 ng/mL~1μg/mL,抗体与对位红和苏丹红I交叉反应率分别为100%和97.8%,与苏丹红II、III、IV和甲苯胺红的交叉反应率很低,与罗丹明类色素无交叉反应。制备的对位红多克隆抗体具有很高的特异性,与其他色素交叉反应率低,可用于对位红在食品中残留的检测。

关 键 词:药敏试验  药敏试验  
收稿时间:2011/1/14 0:00:00
修稿时间:2011/2/28 0:00:00

Synthesis of an Antigen of Para Red and Production of Polyclonal Antibody specific for Para Red
Cheng Mali,Qiu Jing,Liu Hongbin,Yang Shuming,Liang Lin,Zhang Jie,Tang Yu,Shen Hong.Synthesis of an Antigen of Para Red and Production of Polyclonal Antibody specific for Para Red[J].Chinese Agricultural Science Bulletin,2011,27(14):82-86.
Authors:Cheng Mali  Qiu Jing  Liu Hongbin  Yang Shuming  Liang Lin  Zhang Jie  Tang Yu  Shen Hong
Institution:Cheng Mali 1,2,Qiu Jing 2,Liu Hongbin 2,Yang Shuming 2,Liang Lin 1,Zhang Jie 1,Tang Yu 1,Shen Hong 1 ( 1 College of Animal Medicine and Animal Sciences,Beijing University of Agriculture,Beijing 102206,2 Institute of Quality Standard and Test Technology for Agro-product,The Chinese Academy Agricultural Sciences,Beijing 100081)
Abstract:To prepare a polyclonal antibody (pAb) specific for para red, hapten para red was coupled with carrier protein of cationed bovine serum albumin (cBSA) to form a complete antigen by mixed anhydride methods. The conjugates used to immunize Japanese white rabbits were identified with UV scanning spectrum and the specificity of pAb was detected by indirect ELISA and in indirect compete ELISA(icELISA). The couple rate of the antigen was 14:1, the titer of pAb was 1×10 6 and the detection limit in term of 50% inhibition rate was 7.43 ng/mL in the linear range of 0.1 ng/mL-1 μg/mL. The cross-reaction rate between para red and pAb was 100% and was 97.8% between Sudan I and pAb. There was very lower cross-reaction between pAb and para red analogs. The anti-para red polyclonal antibody had a high specificity and was good enough for immunoassay of para red in edible food products.
Keywords:para red  polyclonal antibody (pAb)  ELISA  
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