新型Aβ42寡聚体模拟表位疫苗的制备

发布时间：2018-09-04 09:38

【摘要】：旨在获得一株特异靶向致病性Aβ42寡聚体的模拟表位疫苗。在前期工作中,应用亲和层析的方法从人静脉用免疫球蛋白(Intravenous immune globulin,IVIG)中纯化获得特异性识别Aβ42寡聚体的抗体组分IVIG-AOB。以IVIG-AOB为靶蛋白,应用噬菌体展示技术淘选出一系列Aβ42寡聚体的特异性环状模拟表位多肽,随后将表位多肽融合表达至乙肝病毒核心蛋白(HBcVLP)构建成表位载体疫苗,进而将免疫小鼠,从中筛选出能有效免疫产生抗Aβ42寡聚体抗体的模拟表位疫苗。从噬菌体环形七肽库中筛选出5条特异性结合IVIG-AOB的模拟表位多肽,将其克隆至HBc载体,并在大肠杆菌中成功表达和组装成嵌合表位的HBc-VLP,通过免疫小鼠优选出一株效果最好的Aβ42寡聚体构象表位疫苗HBc-C4。Aβ寡聚体是AD发生发展的主要致病因素。基于Aβ42寡聚体独特的构象表位研制的表位HBc-VLP疫苗诱导机体产生的抗体将能够特异靶向并中和毒性Aβ42寡聚体,而不影响Aβ42的正常生理功能,从而起到从根本上治疗AD的作用。
[Abstract]:The aim of this study was to obtain a mimic epitope vaccine targeting A 尾 42 oligomer. In previous work, IVIG-AOB. was purified from human intravenous immunoglobulin (Intravenous immune globulin,IVIG) by affinity chromatography to identify A 尾 42 oligodeoxynucleotides (A 尾 42). Using IVIG-AOB as target protein, a series of specific cyclic mimic peptides of A 尾 42 oligomer were obtained by phage display technique. Then the epitope peptides were fused into hepatitis B virus core protein (HBcVLP) to construct epitope vector vaccine. Then the mimic epitope vaccine which can effectively immunize mice to produce anti-A 尾 42 oligodeoxymer antibody was screened from the immunized mice. Five mimic peptides with specific binding to IVIG-AOB were screened from phage circular heptapeptide library and cloned into HBc vector. HBc-VLP, which was successfully expressed and assembled into chimeric epitopes in Escherichia coli, was selected as the main pathogenic factor for the development of AD by immunizing mice with the most effective A 尾 42 oligomer conformation epitope vaccine HBc-C4.A 尾 oligomer. The epitope HBc-VLP vaccine based on the unique conformational epitope of A 尾 42 oligomer can induce antibodies to target and neutralize the toxic A 尾 42 oligomer, without affecting the normal physiological function of A 尾 42, and thus play a fundamental role in the treatment of AD.
【作者单位】：齐鲁工业大学生物工程学院;昆明市环境监测中心;中国科学院过程工程研究所;
【基金】：国家自然科学基金项目(81402837) 国家科技重大专项(2014ZX09102041-007)
【分类号】：R749.16

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