William E. Bentley

Bacterial extracellular vesicles (BEVs), including outer membrane vesicles, have emerged as a promising new class of vaccines and therapeutics to treat cancer and inflammatory diseases, among other applications. However, clinical translation of BEVs is hindered by a current lack of scalable and efficient purification methods. Here, we address downstream BEV biomanufacturing limitations by developing a method for orthogonal size‐ and charge‐based BEV enrichment using tangential flow filtration (TFF) in tandem with high performance anion exchange chromatography (HPAEC). The data show that size‐based separation coisolated protein contaminants, whereas size‐based TFF with charged‐based HPAEC dramatically improved purity of BEVs produced by probiotic Gram‐negative Escherichia coli and Gram‐positive lactic acid bacteria (LAB). Escherichia coli BEV purity was quantified using established biochemical markers while improved LAB BEV purity was assessed via observed potentiation of anti‐inflammatory bioactivity. Overall, this work establishes orthogonal TFF + HPAEC as a scalable and efficient method for BEV purification that holds promise for future large‐scale biomanufacturing of therapeutic BEV products.

Quorum sensing, a bacterial process for coordinating community behavior, has inspired scientists to engineer cell‐cell communication for diverse applications. Fundamental knowledge of the molecular underpinnings of quorum sensing systems enabled engineers to rewire quorum sensing circuits in order to alter quorum sensing processes, program control of bacterial populations, and engineer cell‐cell communication. Further, scientific advancements from diverse engineering disciplines have contributed to the design of devices enabling new modes of manipulating or communicating with biological cells. This perspective reviews early and current developments in engineering cell‐cell communication and its applications. Influence of the quorum sensing field on the authors, both engineers, is briefly discussed.