布鲁菌外膜蛋白的糖基化及免疫效果检测  

Immunoassay for targeting vaccine conjugated with glycosylation of Burcella outer membrane proteins

作　　者：付景丽[1] 董炳梅[1] 王若[1] 王家鑫[1] 

机构地区：[1]河北农业大学动物科技学院免疫学实验室,河北保定071000

出　　处：《中国兽医杂志》2011年第4期15-17,共3页Chinese Journal of Veterinary Medicine

基　　金：国家"十一五"科技支撑计划项目(2006BAD04A10);河北省"十一五"科技支撑项目(05240421D);河北省"千万吨奶工程"支撑技术研究(072271462)

摘　　要：为研制安全、有效、新型靶向化布鲁菌外膜蛋白疫苗,采用去污剂连续抽提与电洗脱法分离纯化出布鲁菌外膜蛋白,并进行α-D-甘露吡喃异硫氰酸苯酯糖基修饰、佐剂乳化。依据统计学中完全随机实验设计试验,免疫BALB/c小鼠,收集不同时间点血液样品,ELISA法检测血清中IgG和IFN-γ含量。结果显示分离纯化出37.5 kD与47.2 kD的目的蛋白并成功糖基化修饰;纳米颗粒佐剂乳化后的试验用疫苗免疫小鼠,血清IFN-γ水平呈显著升高趋势(P<0.05);抗体水平略显优势,但差异不显著(P>0.05)。表明试验用疫苗能激发机体的细胞免疫应答,且产生了免疫记忆。本研究为研制布鲁菌新型靶向化疫苗的研发和推广提供了研究依据。In order to design safe,effective and novel targeted Brucella outer membrane protein vaccine,consecutive extraction with eradicator and electric elution was employed to isolate these proteins,glycosylation modified with α-D-mannopyranosyl-phenylisothiocyanate,then emulsified with adjuvant.Based on completely randomized experiments,BALB/c mice were injected,and then samples of serum were collected at different times for the determination of both IgG and IFN-γ levels with ELISA.Results showed that the target proteins of 37.5 kD and 47.2 kD are isolated and glycosylated successfully.After immunizing mice with nanoparticles adjuvant emulsified vaccine,the level of IFN-γ was increased gradually in serum（P0〈.05）.Experimental vaccine has a little advantage in antibody level,but the difference of antibody is not significant（P？0.05）.Experimental vaccine induced cellular immune response with immunological memory.This study paves the way for development and promotion of new Brucella targeted vaccine.

关 键 词：布鲁菌 外膜蛋白 糖基化 佐剂 疫苗 

分 类 号：S852.614[农业科学—基础兽医学]