H1N1型流感病毒神经氨酸酶的原核表达、纯化及免疫原性分析  

Prokaryotic Expression,Purification and Immunogenicity of H1N1 Influenza Virus Neuraminidase

作　　者：杨依丽[1] 唱韶红[1] 巩新[1] 刘波[1] 吴军[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2009年第6期757-760,共4页Letters in Biotechnology

基　　金：国家高技术研究发展计划(2007AA03Z103)

摘　　要：目的:研究H1N1型流感病毒神经氨酸酶(NA)在原核系统中的表达、纯化方法及其免疫原性。方法:构建了大肠杆菌表达载体pET22b-NA,并转化了大肠杆菌BL21(DE3);通过SP-Sepharose Fast Flow柱对重组NA进行分离纯化,并用Sephadex G-25柱对SP柱后获得的NA进行柱上复性;用不同剂量的重组NA免疫BALB/c小鼠,并检测其诱导产生的抗体滴度。结果:大肠杆菌表达的NA以包涵体形式存在,通过分离及柱上复性,纯化得到重组NA;NA抗原的免疫原性是剂量依赖的,随着剂量的增加,其免疫原性相应增强,3次免疫后,3μg NA诱导小鼠产生的抗体滴度最高,为1∶7000。结论:大肠杆菌表达的NA具有一定的诱导小鼠产生针对天然NA的抗体的能力,为流感病毒基因工程疫苗研究提供了初步线索。Objective： To investigate the prokaryotic expression, purification and immunogenicity of H1N1 influenza virus neuraminidase（NA）. Methods： The E.coli expression vector pET＇22b-NA was constructed, and was used to transform to E. coli Bl221 （DE3）. The recombinant NA was separated through SP-Sepharose Fast Flow chromatography and renatured through Sephadex G-25 chromatography. Then, BALB/c mice were immunized with this recombinant NA and antibody titers induced in these mice were tested. Results： NA produced in E.coli was present in the inclusion form. Through sep- aration and renaturation, the purified recombinant NA was harvested. The immunogenicity of NA produced by E.coli was dose-dependent. The immunogenicity of NA was enhanced according to the increasing of the injection dose of NA. After third immunization, the antibody titer elicited by 3 μg NA was 1：7000, it was the highest. Conclusion： NA produced in E.coli could induce nature NA-special antibody titer. This provides clues for the study of influenza genetic engineering vaccine

关 键 词：神经氨酸酶 原核表达 免疫原性 疫苗 H1N1流感病毒 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]