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Study of the stability of proteoliposomes as vehicles for vaccines against Neisseria meningitidis based on recombinant porin complexes

机译:基于重组孔蛋白复合物的蛋白脂质体作为脑膜炎奈瑟菌疫苗载体的稳定性研究

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Although effective against epidemic serogroup B Neisseria meningitidis strains, vaccines based on outer membrane vesicles continue to present important limitations, and great efforts are currently being focused in the development of a variety of new vaccine candidates and in the reformulation of currently existing ones. In this work, three N. meningitidis proteins, the PorA and PorB porins and the RmpM protein, were cloned, purified and incorporated into liposomes to build defined systems. The ability of proteoliposomes to allow the refolding porin complexes, and their stability during storage at 4 ??C and after lyophilization in presence of two cryoprotection agents, glucose and trehalose, were evaluated. This approach allowed to mimic the porin complexes present in natural OMVs, reducing the content of hypervariable protein PorA. During storage at 4 ??C, our systems showed some changes in the morphology and aggregation after three months, while after lyophilization the systems maintained their properties during the whole nine months of storage checked, with glucose allowing the best preservation of the antigenic properties of the proteins in the proteoliposomes. ? 2013 Elsevier B.V.
机译:尽管对付流行性脑膜炎奈瑟氏菌脑膜炎奈瑟氏球菌菌株有效,但基于外膜囊泡的疫苗仍然存在重要的局限性,并且目前正在集中精力开发各种新的候选疫苗和重新配制现有的候选疫苗。在这项工作中,克隆,纯化了三种脑膜炎奈瑟氏球菌蛋白,PorA和PorB孔蛋白以及RmpM蛋白,并将其掺入脂质体中以构建确定的系统。评价了蛋白脂质体允许再折叠孔蛋白复合物的能力,以及它们在4℃下储存期间以及在两种冷冻保护剂(葡萄糖和海藻糖)存在下冻干后的稳定性。这种方法可以模拟天然OMV中存在的孔蛋白复合物,从而降低高变蛋白PorA的含量。在4°C下储存期间,我们的系统在三个月后显示出一些形态和聚集的变化,而冻干后,系统在整个九个月的储存过程中都保持了其特性,而葡萄糖可以最佳地保留其抗原性。蛋白脂质体中的蛋白质。 ? 2013 Elsevier B.V.

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