基于新载体的b型流感嗜血杆菌结合疫苗的制备及免疫原性研究  

Preparation and immunogenicity study of the Haemophilus influenzae type b conjugate vaccine based on a novel carrier protein

作　　者：苗丹青 李军强 杨鸣鸣 邵忠琦 朱涛[1] 

机构地区：[1]天津科技大学,天津300457 [2]天津康希诺生物技术有限公司,天津300457

出　　处：《中国新药杂志》2016年第7期739-744,共6页Chinese Journal of New Drugs

基　　金：国家"重大新药创制"科技重大专项(2015ZX09102009)

摘　　要：目的:评价Hin47-HiD融合蛋白作为b型流感嗜血杆菌结合疫苗新型载体的可行性。方法:应用重叠延伸PCR法构建Hin47-HiD融合基因,克隆至表达载体p ET-9a,并转化至大肠杆菌中诱导表达。融合蛋白经柱层析纯化后与Hib多糖结合,免疫小鼠并用间接ELISA法测定小鼠血清中针对Hib多糖和Hin47、HiD蛋白的抗体滴度。结果:本研究表达、纯化了Hin47-HiD融合蛋白,并制备了以该蛋白为载体的Hib结合疫苗;两针免疫后,免疫Hib-Hin47-HiD结合物的小鼠血清中的抗Hib抗体水平远高于接种Hib多糖的小鼠;此外,Hib-Hin47-HiD结合物还诱导出高水平的针对Hin47和HiD的抗体。结论:Hin47-HiD融合蛋白可以有效的提高Hib多糖的免疫原性,是一个有潜力的新型结合疫苗蛋白载体。Objective： To evaluate the feasibility of the fusion protein（ Hin47-HiD） as a new carrier protein for the preparation of the Haemophilus influenzae type b（ Hib） conjugate vaccine. Methods： A DNA fragment coding for the Hin47-HiD fusion protein was constructed by overlap PCR and cloned into pET-9a vector for expression in the E. coli host cell BL21（ DE3）. The fusion protein was purified by column chromatography and used for the preparation of the Hib polysaccharide conjugate vaccine. BALB / c mice were immunized with the Hib-Hin47-HiD conjugate,and antibody titers in mouse sera against the Hib polysaccharide as well as the carrier protein were analyzed using indirect ELISA. Results： The Haemophilus influenzae type b conjugate vaccine prepared using the fusion protein was highly immunogenic,producing high antibody levels in mouse sera against the Hib polysaccharide following two injections. In addition,the Hib-Hin47-HiD conjugate also generated good antibody responses against both Hin47 and HiD. Conclusion： The Hin47-HiD fusion protein can effectively improve the immunogenicity of the Hib polysaccharide,and may be used as an alternative carrier protein for the preparation of the conjugate vaccine.

关 键 词：b型流感嗜血杆菌荚膜多糖 结合疫苗 融合蛋白 免疫原性 

分 类 号：R86[医药卫生—临床医学] R979.5