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。
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%.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第5期181-185,共5页
Food Science
基金
浙江省重点科技创新团队项目(2010R50028)
浙江省科技成果转化项目(2011E61018)
浙江省重大科技专项重点项目(2012C12013-3)
