A群脑膜炎球菌多糖-重组蛋白Pep10结合物的制备及其免疫原性  

Preparation and Immunogenicity of Group A Meningococcal Polysaccharide-Recombinant Protein Pep10 Conjugate

作　　者：刘汉平[1] 薛红刚[1] 方志正[1] 

机构地区：[1]武汉生物制品研究所基因工程室,武汉430060

出　　处：《中国生物制品学杂志》2008年第7期591-594,共4页Chinese Journal of Biologicals

摘　　要：目的制备A群脑膜炎球菌多糖(GAMP)-重组蛋白Pep10结合物,并检测其免疫原性,探讨重组蛋白Pep10作为一种候选载体蛋白的可行性。方法GAMP经溴化氰(CNBr)活化后,共价接合己二酰肼(ADH)手臂,在碳二亚胺(EDAC)催化下与重组蛋白Pep10偶联,制备结合物GAMP-ADH-Pep10。纯化后免疫BALB/c小鼠,用间接ELISA法检测小鼠血清中抗GAMP IgG抗体水平,并分析载体诱导的免疫抑制率。结果所制备的多糖衍生物GAMP-ADH及多糖-蛋白结合物GAMP-ADH-Pep10均具有GAMP抗原特异活性,免疫小鼠后可诱生比多糖单独免疫更高水平的GAMP血清IgG抗体,且接近GAMP-ADH-TT疫苗诱生的抗体水平,并能形成免疫记忆。重组蛋白Pep10诱导的免疫抑制率远低于TT诱导的免疫抑制率。结论制备的重组蛋白Pep10与A群脑膜炎球菌多糖偶联能发挥较强的载体效应,为研究理想载体蛋白提供了新的思路。Objective To prepare group A meningococcal polysaccharide （GAMP） recombinant protein Pep10 conjugate, study its immunogenicity and explore the feasibility of the conjugate as a candidate carrier protein. Methods GAMP was activated with cyanogen bromide and combined with 1, 6-adipic acid dihydrazide （ADH）. The prepared polysaccharide derivative GAMP-ADH was coupled to recombinant protein PeplO with N-（3-dimethylaminopropyl）-N′-ethylcarbodiimide hydrochloride （EDAC）. Immunize BALB/c mice with the produced GAMP-ADH-Pep10 conjugate after purification, determine the serum IgG level against GAMP by indirect ELISA, and analyze the immune suppression induced by carrier protein. Results Both GAMP-ADH and GAMP-ADH-Pep10 showed GAMP antigenic specificity. The serum IgG level against GAMP induced by GAMP-ADH-Pep10 conjugate was significantly higher than that by GAMP and showed no significant difference with that by GAMP-ADH-TT vaccine. The conjugate also induced immunological memory. However, the immune suppression rate induced by recombinant protein Pep10 was significantly lower than that by TT. Conclusion The recombinant protein Pep10 conjugated with GAMP showed strong carrier effect, which provided a novel route for the development of ideal carrier protein.

关 键 词：A群脑膜炎球菌多糖 载体蛋白 结合物 免疫原性 

分 类 号：R373[医药卫生—病原生物学] R378.15[医药卫生—基础医学]