1. PhagoScreener: A novel phagogram platform based on a capillary-wave microbioreactor.
- Author
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Viebrock, Kevin, Wilhelm, Jana, Rölke, Bea, Pastwa, Leon, Schrader, Selina M., Meinen, Sven, Dietzel, Andreas, Dohnt, Katrin, Ziehr, Holger, Korf, Imke H.E., Bohle, Kathrin, and Krull, Rainer
- Subjects
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ESCHERICHIA coli , *ANTIBIOTIC overuse , *HYDROPHILIC surfaces , *MICROPLATES , *AGAR plates , *BACTERIOPHAGES - Abstract
Due to the overuse of antibiotics, the number of multidrug-resistant pathogen bacteria is rising in recent years posing a serious threat to human health. One promising alternative for treatment is the application of phage therapy using highly selective bacteriophages. Because of their selectivity, individual screens called phagograms for each patient are required to select phages from a phage library. Phagograms are mostly performed via bacterial cultivation on double layer agar plates and phage addition causing bacterial lysis. However, these assays are work-intensive and have a low ability for parallelization and automation. Hence, highly parallelizable and automatable microbioreactors in the lowest microliter scale could offer an economic solution increasing the throughput of phagograms. This paper demonstrates the applicability of a novel capillary-wave microbioreactor (cwMBR) to perform phagograms. Due to its small volume of only 7 µL and the open-droplet design, it can be easily automated and parallelized in future. Furthermore, the ability of online biomass measurement makes the cwMBR a perfect phagogram platform in the future. Herein, phagograms with E. coli and different concentrations of the phages MM02 and EASG3 were performed as proof of concept for phagograms in the cwMBR. Thereby, the cwMBR was able to measure differences in lysis kinetics of different phages. Furthermore, the phagograms were compared to those in conventional microtiter plate readers revealing the cwMBR as ideal alternative for phagograms as it combines favorable mixing conditions and a phage repellent hydrophilic glass surface with online biomass measurement in an open-droplet design for future parallelization and automation. [Display omitted] • Phage sensitivity tests called phagograms are mandatory for phage therapy. • State of the art phagograms are not highly parallelizable and automatable. • Novel capillary-wave microbioreactor (cwMBR) can perform parallelized phagograms. • cwMBR combines parallelizability and automatability with online biomass measurement. • cwMBR is able to measure lysis kinetics of phages. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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