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Spontaneously released Neisseria meningitidis outer membrane vesicles as vaccine platform: production and purification

机译：自发地释放的脑膜炎脑膜外膜囊泡作为疫苗平台：生产和纯化

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Outer membrane vesicles (OMVs) are nanoparticles produced by Gram-negative bacteria that can be used as vaccines. The application of OMVs as vaccine component can be expanded by expressing heterologous antigens on OMVs, creating an OMV-based antigen presenting platform. This study aims to develop a production process for such OMV-based vaccines and studies a production method based on meningococcal OMVs that express heterologous antigens on their surface. As a proof of concept, the Borrelia burgdorferi antigens OspA and OspC were expressed on Neisseria meningitidis OMVs to create a concept anti-Lyme disease vaccine. Production of OMVs released in the culture supernatant was induced by high dissolved oxygen concentrations and purification was based on scalable unit operations. A crude recovery of 90 mg OMV protein could be obtained per liter culture. Expressing heterologous antigens on the OMVs did result in minor reduction of bacterial growth, while OMV production remained constant. The antigen expression did not alter the OMV characteristics. This study shows that production of well characterized OMVs containing heterologous antigens is possible with high yields by combining high oxygen concentrations with an optimized purification process. It is concluded that heterologous OMVs show potential as a vaccine platform. (C) 2019 The Authors. Published by Elsevier Ltd.
机译：外膜囊泡（OMV）是由革兰氏阴性细菌产生的纳米颗粒，其可用作疫苗。通过在OMV上表达异源抗原，可以通过在OMV上表达异源抗原来扩增OMV作为疫苗组分的应用，从而产生基于OMV的抗原呈现平台。本研究旨在开发基于OMV的疫苗的生产过程，并研究基于脑膜炎球菌OMV的生产方法，其表达其表面上的异源抗原。作为概念证明，Borrelia Burgdorferi抗原OSPA和OSPC在Neisseria Meningitidis OMV中表达，以创建一个概念抗莱姆病疫苗。通过高溶解的氧浓度诱导培养上清液中释放的OMV的生产，并纯化基于可扩展单元操作。每升培养物可以获得90mg omv蛋白的粗产回。在OMV上表达异源抗原确实导致细菌生长的轻微降低，而OMV产生仍然是恒定的。抗原表达没有改变OMV特征。该研究表明，通过将高氧浓度与优化的纯化过程组合，可以高产率的含有异源抗原的良好表征OMV的产生。结论是，异源OMVs显示出作为疫苗平台的潜力。 （c）2019年作者。 elsevier有限公司出版

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