NGAL原核表达及多克隆抗体的制备  

Prokaryotic expression and preparation of polyclonal antibody of NGAL

作　　者：王小中[1] 熊火梅[1] 黄波[1] 李静[2] 肖芸[1] 章海斌[1] 刘静[1] 王彩纹[3] 

机构地区：[1]南昌大学第二附属医院检验科,330006 [2]南昌大学第一附属医院检验科,330006 [3]江西省胸科医院检验科,南昌330006

出　　处：《国际检验医学杂志》2011年第16期1789-1791,共3页International Journal of Laboratory Medicine

基　　金：江西省科技厅科技支撑项目(赣科发计字[2007]196号);江西省教育厅科技项目(赣教技字[2007]100号)

摘　　要：目的构建人中性粒细胞明胶酶相关脂质运载蛋白(NGAL)的原核表达载体,制备兔抗人NGAL多克隆抗体。方法利用DNA重组技术构建原核表达载体pET28a-NGAL,诱导表达NGAL融合蛋白,分离该融合蛋白后,注射入免疫家兔制备多克隆抗体。结果成功获得NGAL,免疫动物所得抗血清效价为1∶4000,该抗体能够识别原核表达的NGAL。结论成功克隆NGAL基因并制备了特异性的兔抗人NGAL多克隆抗体。Objective To construct the prokaryotic expression vector pET28a-NGAL and prepare the specific polyclonal antibody against NGAL.Methods Recombinant plasmid pET28a-NGAL was expressed in E.coli after induction with IPTG. The NGAL fusion protein was identified by sodium dodecyl sulfate-polymacrylamide gel electrophoresis,and was extracted and injected into rabbits to produce polyclonal antibody.Results The protein of NGAL was obtained by recombination expression. The titers of the antiserum were up to 1∶4 000.This antibody could specifically recognize the NGAL protein expressed in the E.coli system.Conclusion The gene encoding NGAL is successfully cloned and the specific polyclonal antibody against NGAL was prepared.

关 键 词：基因 遗传 融合表达 多克隆抗体 人中性粒细胞明酶相关脂质运载蛋白 

分 类 号：R392[医药卫生—免疫学]