Bacterial membrane vesicles (BMVs) are bi-layered nanostructures derived from Gram-negative and Gram-positive bacteria. Among other pathophysiological roles, BMVs are critical messengers in intercellular communication. As a result, BMVs are emerging as a promising technology for the development of numerous therapeutic applications. Despite the remarkable progress in unveiling BMV biology and functions in recent years, their successful isolation and purification have been limited. Several challenges related to vesicle purity, yield, and scalability severely hamper the further development of BMVs for biotechnology and clinical applications. This review focuses on the current technologies and methodologies used in BMV production and purification, such as ultracentrifugation, density-gradient centrifugation, size-exclusion chromatography, ultrafiltration, and precipitation. We also discuss the current challenges related to BMV isolation, large-scale production, storage, and stability that limit their application. More importantly, the present work explains the most recent strategies proposed for overcoming those challenges. Finally, we summarize the ongoing applications of BMVs in the biotechnological field.

中文翻译：

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用于生物技术应用的细菌膜囊泡的生产和纯化：挑战和机遇

细菌膜囊泡 (BMV) 是源自革兰氏阴性和革兰氏阳性细菌的双层纳米结构。在其他病理生理作用中，BMV 是细胞间通讯的关键信使。因此，BMV 正在成为开发众多治疗应用的有前途的技术。尽管近年来在揭示 BMV 生物学和功能方面取得了显着进展，但它们的成功分离和纯化受到限制。与囊泡纯度、产量和可扩展性相关的若干挑战严重阻碍了 BMV 在生物技术和临床应用中的进一步发展。本综述侧重于 BMV 生产和纯化中使用的当前技术和方法，例如超速离心、密度梯度离心、体积排阻层析、超滤、沉淀。我们还讨论了当前限制其应用的与 BMV 隔离、大规模生产、存储和稳定性相关的挑战。更重要的是，目前的工作解释了为克服这些挑战而提出的最新策略。最后，我们总结了 BMV 在生物技术领域的持续应用。