辣根过氧化物酶标记兔抗丝状噬菌体衣壳蛋白抗体的制备与初步应用  

Preparation and preliminary application of the rabbit antibody against capsid protein of the filamentous phage labeled with horseradish peroxidase

作　　者：徐晶晶[1] 李迪[1] 曹永生[1] 高明春[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030

出　　处：《黑龙江畜牧兽医》2013年第3期114-116,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金："十二五"国家科技支撑计划项目(2012BAD12B05;2012BAD12B03);现代农业(奶牛)产业技术体系项目(CARS-37);黑龙江省科技攻关项目(GA09B302)

摘　　要：为了制备用于噬菌体展示肽库的筛选及鉴定等环节的辣根过氧化物酶(HRP)标记的兔抗丝状噬菌体(M13)衣壳蛋白抗体,试验制备了兔源的M13KE多克隆抗体,将多克隆抗体经饱和硫酸铵法和Protein G Resin层析柱纯化后运用过碘酸钠法对其进行辣根过氧化物酶标记,对标记前和标记后纯化产物进行活性测定,同时对标记后产物进行Western-blot鉴定,以及将其应用到验证已知的阳性噬菌体H11、H12、H14、H16、H18中。结果表明:兔抗M13KE多克隆抗体效价高于1∶32 000,标记后活性高于1∶4 000,标记抗体与已知阳性噬菌体的ELISA鉴定结果与前期测序结果一致。说明此酶标抗体有一定的应用价值。To prepare rabbit antibody against M13 capsid protein labeled with horseradish peroxidase for the screening and identification of phage display peptide library, the rabbit anti - M13 KE virus polyclonal antibody was prepared, and then was labeled with HRP by sodium iodide after precipitated and purified in saturated solution of sulfate of ammonia and Protein G Resin purification. The activity of purified product was determined before and after labeled with HRP, and the labeled product was determined by western blot, and then was applied into the verification of known positive phage H11, H12, H14, H16 and H18. The results showed that the rabbit anti - M13 polyclonal antibody titer was higher than 1：32 000 and the activity of the labeled product was higher than 1：4 000. The results of ELISA identification between enzyme - labeled antibody and known positive phages were consistent with the previous sequencing results. It indicates that the enzyme - labeled antibody has certain application value.

关 键 词：M13KE噬菌体 酶标抗体 辣根过氧化物酶(HRP) 酶联免疫吸附试验(ELISA) 

分 类 号：S852.4[农业科学—基础兽医学]