消减免疫法制备氨苄青霉素抗体  

PREPARATION OF AMPICILLIN ANTIBODY BY SUBTRACTIVE IMMUNIZATION

作　　者：王彩红[1] 宁保安[1] 王红勇[1] 李晓莉[1] 刘晶 连璐 高志贤[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所,天津300050 [2]天津环境与食品安全风险监控技术重点实验室

出　　处：《解放军预防医学杂志》2010年第2期90-94,共5页Journal of Preventive Medicine of Chinese People's Liberation Army

基　　金：天津市应用基础及前沿技术研究计划(No.OPJCZDJC17700)

摘　　要：目的应用消减免疫法制备氨苄青霉素(Amp)多克隆抗体,为动物性食品安全快速检测及进出口贸易提供技术支持。方法以生物偶联法制备Amp-OVA与Amp-BSA,以紫外分光光度法与SDS-PAGE鉴定完全抗原,以环磷酰胺药物消减免疫,以ELISA检测免疫耐受情况;直接ELISA法检测抗体效价,竞争ELISA检测抗体特异性与灵敏度并评价消减免疫效果。结果成功合成了Amp-OVA完全抗原,消减免疫使小鼠获得了对OVA的耐受,再经多次Amp-OVA免疫后,与未消减组小鼠(血清中针对OVA的效价1:10 000)相比,消减组小鼠血清针对OVA的抗体效价(1:1 000)明显降低。结论相对于常规免疫法,采用消减免疫法更易获得针对Amp的多克隆抗体。Objective To obtain ampicillin（Amp） polyclonal antibody by subtractive immunization,so as to provide technical support for rapid detection of food safety and import-export trade of animal food.Methods Ampicillin was coupled to BSA and OVA.The results were indicated by ultraviolet spectrophotometry and SDS-PAGE.Subtractive immunization was performed by cyclophosphamide.The tolerance of OVA was dectected by ELISA.Direct ELISA and competitive ELISA were used to characterize the antibody.Results The antigen of Amp-OVA was compounded successfully.The tolerance of OVA was obtained by subtractive immunization.The titre of subtractive immunization group（1∶1000） was obviously lower than that of unsubtractive immunization group（1∶10 000）.Conclusion Ampicillin polyclonal antibody can be obtained more easily by subtractive immunization than common unsubtractive immunization.

关 键 词：消减免疫 环磷酰胺 氨苄青霉素 抗体 

分 类 号：R446.6[医药卫生—诊断学]