Monolith Based Chromatography Is a Suitable Tool to Prepare and Analyze E. coli – Outer Membrane Vesicles

Intranasal administration of Escherichia coli O83 – outer membrane vesicles (OMVs) was found to reduce allergic airway inflammation induced by ovalbumin in mice. Production of sufficient amounts of OMVs to supply further clinical trials implies the need for scalable methods and precise analytics. Different orthogonal methods are required to provide a complete picture of the process. We present an exemplary OMV preparation, demonstrating robustness of our chromatographic platform. As shown before for various types of extracellular vesicles from animal and human cell origin, the platform was proved suitable also to isolate OMV´s produced by E. coli. The purification process was monitored using three common analytical techniques – nanoparticle tracking analysis, flow cytometry and PATFix chromatography. The results demonstrate that chromatographic preparative methods are suitable to produce OMVs and that orthogonal analytical methods are mandatory to paint an accurate picture through the whole preparative process.

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