Abstract
Due to its unique capability to analyze a large number of single cells for several parameters simultaneously, flow cytometry has changed our understanding of the behavior of cells in culture and of the population dynamics even of clonal populations. The potential of this method for biotechnological research, which is based on populations of living cells, was soon appreciated. Sorting applications, however, are still less frequent than one would expect with regard to their potential. This review highlights important contributions where flow cytometric cell sorting was used for physiological research, protein engineering, cell engineering, specifically emphasizing selection of overproducing cell lines. Finally conclusions are drawn concerning the impact of cell sorting on inverse metabolic engineering and systems biology.
Highlights
The establishment of flow cytometry as the first single cell analysis method with the potential to describe the distribution of cellular properties within a large number of cells has considerably changed our knowledge of cell populations
While cell sorting is still mainly applied for clinical purposes, there is an increasing interest in biotechnology to utilize its potential for library screening or strain development
The aim of this review was to structure the fields of cell sorting applications in biotechnology, and to highlight many of the examples published over the last years
Summary
The establishment of flow cytometry as the first single cell analysis method with the potential to describe the distribution of cellular properties within a large number of cells has considerably changed our knowledge of cell populations. Analysis of cells in culture has shown that with the exception of DNA content, all other cellular components are distributed over a wide range, which is the reason why such parameters are usually presented on a logarithmic scale in flow cytometry histograms This variation of cellular properties is of special interest for strain improvement purposes, as it allows the sorting of cells with diverging and potentially optimized properties. With flow cytometry, analyzing 104 cells is standard procedure and it is possible to look at millions of cells without much trouble This feature is especially important for biotechnological applications, because it is usually the rare cell which is of interest: the cell with altered properties, a higher production rate, better metabolic parameters or containing the protein with a higher binding affinity. Yeasts antibody surface display peptide surface display intra- and extracellular enzymes yeast hybridization, library cloning bacteria acid tolerance high cell density, low growth rate protein alkaloids FITC/antibiotic production
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