title: Flow cytometric analysis of E. coli on agar plates: implications for recombinant protein production.
abstract: Recombinant protein production in bacterial hosts is a commercially important process in the pharmaceutical industry. Optimisation of such processes is of critical importance for process productivity and reproducibility. Here, flow cytometry methods were developed to assess characteristics of bacteria during two process steps that are infrequently studied: agar plate culture and liquid culture set-up. During storage on agar plates, three discrete populations of varying green fluorescence intensity were observed along with a progressive shift of cells from the high green fluorescence population to an intermediate green fluorescence population, observed to be due formation of amyloid inclusion bodies. The dynamics of cellular fluorescence and scatter properties upon setup of liquid cultures were also assessed. These methods have the potential to improve the development of fermentation set-up, a currently little-understood area.
authors.affiliation: Bioengineering, School of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK, chrisjwyre@gmail.com.
authors.forename: Chris
authors.initials: C
authors.lastname: Wyre
authors.forename: Tim W
authors.initials: TW
authors.lastname: Overton
mesh_terms: D000362:Agar; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D000362:Agar