School of Biosciences, Firth Court, University of Sheffield, Sheffield, United Kingdom.
GjerdeTechnologies, Saratoga, California, USA.
Microbiol Spectr. 2022 Oct 26;10(5):e0245022. doi: 10.1128/spectrum.02450-22. Epub 2022 Sep 21.
The isolation of pure, single colonies lies at the heart of experimental microbiology. However, a microbial colony typically contains around 1 million cells at all stages of the life cycle. Here, we describe a novel cell chromatography method that facilitates the capture, purification, and interrogation of microbial cell populations from both single and mixed cultures. The method described relies on, but is not limited to, differences in surface charge to separate bacterial strains. The method is fully biocompatible, leading to no significant loss of cell viability. The chromatographic capture of cells, combined with selective elution methods, facilitates a greater level of experimental control over the sample inputs required for downstream high-throughput and high-sensitivity analytical methods. The application of the method for interrogating the antibiotic resistance of bacterial strains and for the separation of bacteria from environmental samples is illustrated. This is the first report of a method for separating microbial cells using chromatography, with full retention of cell viability. Differences in the surface chemistry of microbial cells provides a means of attracting cells to immobilized microbeads. Some cells are attracted, and some are repelled. The differences in, for example, surface charge can be harnessed to capture, interrogate, and separate environmental samples, thus circumventing the need to use conventional bacterial plating methods. This method will greatly facilitate drug discovery and bioprospecting for novel microbial compounds.
纯单菌落的分离是实验微生物学的核心。然而,一个微生物菌落通常包含处于生命周期各个阶段的约 100 万个细胞。在这里,我们描述了一种新颖的细胞色谱法,该方法有助于从单一和混合培养物中捕获、纯化和检测微生物细胞群体。所描述的方法依赖于但不限于表面电荷的差异来分离细菌菌株。该方法完全生物相容,不会导致细胞活力显著丧失。细胞的色谱捕获,结合选择性洗脱方法,为下游高通量和高灵敏度分析方法所需的样本输入提供了更大程度的实验控制。该方法在检测细菌菌株的抗生素耐药性和从环境样本中分离细菌方面的应用进行了说明。这是首次报道使用色谱法分离微生物细胞的方法,同时保持细胞活力的完整。微生物细胞表面化学性质的差异提供了一种吸引细胞固定化微球的方法。有些细胞被吸引,有些则被排斥。例如,表面电荷的差异可以被利用来捕获、检测和分离环境样本,从而避免使用传统的细菌平板方法。这种方法将极大地促进药物发现和新型微生物化合物的生物勘探。