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吞噬体筛选器:基于毛细波微生物反应器的新型吞噬体平台。

PhagoScreener: A novel phagogram platform based on a capillary-wave microbioreactor.

机构信息

Institute of Biochemical Engineering, Technische Universität Braunschweig, Rebenring 56, 38106 Braunschweig, Germany; Center of Pharmaceutical Engineering, Technische Universität Braunschweig, Franz-Liszt-Str. 35a, 38106 Braunschweig, Germany.

Fraunhofer Institute for Toxicology and Experimental Medicine, Inhoffenstr. 7, 38124 Braunschweig, Germany; Center of Pharmaceutical Engineering, Technische Universität Braunschweig, Franz-Liszt-Str. 35a, 38106 Braunschweig, Germany.

出版信息

N Biotechnol. 2024 Nov 25;83:188-196. doi: 10.1016/j.nbt.2024.08.502. Epub 2024 Aug 22.

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.

摘要

由于抗生素的过度使用,近年来多药耐药病原体细菌的数量不断增加,对人类健康构成了严重威胁。一种有前途的治疗替代品是应用噬菌体疗法,使用高度选择性的噬菌体。由于其选择性,需要为每个患者进行单独的筛选,称为噬菌体图谱,以从噬菌体文库中选择噬菌体。噬菌体图谱大多通过双层琼脂平板上的细菌培养和添加噬菌体导致细菌裂解来进行。然而,这些测定方法劳动强度大,并行化和自动化能力低。因此,在最低微升规模下具有高度可并行化和自动化的微生物反应器可以提供一种经济的解决方案,提高噬菌体图谱的通量。本文展示了一种新型毛细管波微生物反应器 (cwMBR) 在执行噬菌体图谱方面的适用性。由于其体积仅为 7 µL,且采用开放式液滴设计,因此将来可以轻松实现自动化和并行化。此外,在线生物量测量的能力使 cwMBR 成为未来噬菌体图谱的完美平台。在此,使用大肠杆菌和不同浓度的噬菌体 MM02 和 EASG3 进行噬菌体图谱,作为 cwMBR 中噬菌体图谱的概念验证。因此,cwMBR 能够测量不同噬菌体的裂解动力学差异。此外,将噬菌体图谱与传统微孔板阅读器进行比较,表明 cwMBR 是噬菌体图谱的理想替代品,因为它结合了有利的混合条件和抗噬菌体的亲水玻璃表面,以及在开放式液滴设计中进行在线生物量测量,以实现未来的并行化和自动化。

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