Institute for Translational Vaccinology (Intravacc), Process Development Bacterial Vaccines, P.O. Box 450, 3720, AL, Bilthoven, The Netherlands.
Bioprocess Engineering, Wageningen University, P.O. Box 16, 6700, AA, Wageningen, The Netherlands.
Appl Microbiol Biotechnol. 2019 Dec;103(23-24):9401-9410. doi: 10.1007/s00253-019-10163-z. Epub 2019 Nov 1.
Outer membrane vesicles (OMVs) are nanoparticles secreted by Gram-negative bacteria that can be used for diverse biotechnological applications. Interesting applications have been developed, where OMVs are the basis of drug delivery, enzyme carriers, adjuvants, and vaccines. Historically, OMV research has mainly focused on vaccines. Therefore, current OMV production processes have been based on batch processes. The production of OMVs in batch mode is characterized by relatively low yields and high costs. Transition of OMV production processes from batch to continuous processes could increase the volumetric productivity, reduce the production and capital costs, and result in a higher quality product. Here, we study the continuous production of Neisseria meningitidis OMVs to improve volumetric productivity. Continuous cultivation of N. meningitidis resulted in a steady state with similar high OMV concentrations as are reached in current batch processes. The steady state was reproducible and could be maintained for at least 600 h. The volumetric productivity of a continuous culture reached 4.0 × 10 OMVs per liter culture per day, based on a dilution rate of 1/day. The tested characteristics of the OMVs did not change during the experiments showing feasibility of a continuous production process for the production of OMVs for any application.
外膜囊泡（OMVs）是革兰氏阴性菌分泌的纳米颗粒，可用于多种生物技术应用。已经开发出了有趣的应用，其中 OMV 是药物输送、酶载体、佐剂和疫苗的基础。从历史上看，OMV 的研究主要集中在疫苗上。因此，目前的 OMV 生产工艺主要基于分批工艺。在分批模式下生产 OMV 的特点是产率相对较低，成本较高。将 OMV 生产工艺从分批向连续工艺过渡可以提高体积生产率，降低生产成本和资本成本，并生产出更高质量的产品。在这里，我们研究了脑膜炎奈瑟菌 OMV 的连续生产，以提高体积生产率。脑膜炎奈瑟菌的连续培养导致了类似于当前分批工艺中达到的高 OMV 浓度的稳定状态。该稳定状态具有重现性，至少可以维持 600 小时。基于 1/天的稀释率，连续培养的体积生产率达到了每升培养物每天 4.0×10 OMVs。在实验过程中，OMVs 的测试特性没有发生变化，表明连续生产工艺对于任何应用的 OMV 生产都是可行的。
