链霉素人工抗原及多克隆抗体的制备与鉴定

doi:10.7506/spkx1002-6630-201305038

食品科学 ›› 2013, Vol. 34 ›› Issue (5): 181-185.doi: 10.7506/spkx1002-6630-201305038

链霉素人工抗原及多克隆抗体的制备与鉴定

奚茜¹,李沐洁¹,龚云飞¹,陈宗伦²,方结红¹,王唯芬¹,张明洲^1*

1. 中国计量学院
2. 中国计量学院生命科学学院

收稿日期:2011-12-18 修回日期:2013-01-21 出版日期:2013-03-15 发布日期:2013-04-16
通讯作者: 张明洲 E-mail:zmzcjlu@cjlu.edu.cn
基金资助:
浙江省重大科技专项;浙江省重点科技创新团队

Preparation and Identification of Streptomycin Artificial Antigen and Its Polyclonal Antibody

XI Xi1，LI Mu-jie1，GONG Yun-fei1，CHEN Zong-lun2，FANG Jie-hong1，WANG Wei-fen1，ZHANG Ming-zhou1,*

1. Zhejiang Provincial Key Laboratory of Biometrology, Inspection and Quarantine, College of Life Science, China Jiliang University,
Hangzhou 310018, China；2. Hangzhou DNA Scitech Co. Ltd., Hangzhou 310013, China

Received:2011-12-18 Revised:2013-01-21 Online:2013-03-15 Published:2013-04-16
Contact: ZHANG Ming-zhou E-mail:zmzcjlu@cjlu.edu.cn

摘要/Abstract

摘要：

通过研究链霉素(SM)人工抗原的合成和多克隆抗体的制备，建立链霉素残留的免疫分析方法。以氧-缩甲基羟胺肟化链霉素的醛基，引入羧基，再用碳二亚胺法将半抗原与蛋白质载体连接，合成免疫原SM-cBSA。用戊二醛法合成包被原SM-OVA。用紫外扫描、SDS-PAGE分析偶联情况，计算得链霉素与cBSA、OVA的分子偶联比分别为7.6:1与17.7:1。经动物免疫获得效价为1:40000的链霉素多克隆抗体，采用间接竞争ELISA测定IC50为3.32ng/mL，与双氢链霉素的交叉反应率为105.21%，与同类的其他药物均无交叉反应。

关键词: 链霉素, 人工抗原, 多克隆抗体, 间接ELISA法

Abstract:

In order to develop an immunoassay for treptomycin (SM) residues, the synthesis of antigen and preparation of
polyclonal antibody for SM were explored. In this study, immunogen SM-cBSA was synthesized by EDC after inserting
carboxy group by O-carboxymethyl-hydroxylamine. Coating antigen SM-OVA was synthesized by glutaraldehyde.
Both SM-cBSA and SM-OVA were identified with UV-visible spectroscopy and SDS-PAGE to reveal the successful
coupling with a molar coupling ratio of 7.6:1 and 17.7:1, respectively. During animal immunization, the antibody
titer and IC50 were detected by indirect ELISA and measured by competitive ELISA to be 1:40000 and 3.32 ng/mL,
respectively. Meanwhile, the cross-reactivity of streptomycin polyclonal antibody with dihydrostreptomycin was 105.21%.

Key words: streptomycin, artificial antigen, polyclonal antibody, indirect ELISA

中图分类号:

S859.84

奚茜李沐洁龚云飞陈宗伦方结红王唯芬张明洲. 链霉素人工抗原及多克隆抗体的制备与鉴定[J]. 食品科学, 2013, 34(5): 181-185.

XI Xi1，LI Mu-jie1，GONG Yun-fei1，CHEN Zong-lun2，FANG Jie-hong1，WANG Wei-fen1，ZHANG Ming-zhou1,*. Preparation and Identification of Streptomycin Artificial Antigen and Its Polyclonal Antibody[J]. FOOD SCIENCE, 2013, 34(5): 181-185.

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链霉素人工抗原及多克隆抗体的制备与鉴定

Preparation and Identification of Streptomycin Artificial Antigen and Its Polyclonal Antibody

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