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Monitoring and quantification of inclusion body formation in Escherichia coli by multi-parameter flow cytometry
Royal Institute of Technology, Sweden.
Royal Institute of Technology, Sweden.
Mälardalen University, Department of Biology and Chemical Engineering.
University of Birmingham, UK.
Show others and affiliations
2005 (English)In: Biotechnology Letters, ISSN 0141-5492, Vol. 27, no 13, 919-926 p.Article in journal (Refereed) Published
Abstract [en]

Multi-parameter flow cytometry was used to monitor the formation of promegapoietin (PMP) inclusion bodies during a high cell density Escherichia coli fed-batch fermentation process. Inclusion bodies were labelled with a primary antibody and then with a secondary fluorescent antibody. Using this method it was possible to detect PMP inclusion body formation with a high specificity and it was possible to monitor the increased accumulation of the protein with process time (6-48 mg PMP/g CDW) whilst highlighting population heterogeneity.

Place, publisher, year, edition, pages
2005. Vol. 27, no 13, 919-926 p.
Keyword [en]
Bioreactors/microbiology, Chromatography; High Pressure Liquid, Escherichia coli/growth & development/*metabolism, Fermentation, Flow Cytometry/*methods, Fluorescent Dyes/chemistry, Humans, Inclusion Bodies/chemistry/*metabolism, Interleukin-3, Microscopy; Fluorescence, Receptors; Interleukin-3/chemistry/genetics/metabolism, Recombinant Fusion Proteins, Recombinant Proteins/metabolism, Thrombopoietin/chemistry/genetics/metabolism
National Category
Biophysics
Identifiers
URN: urn:nbn:se:mdh:diva-2154DOI: 10.1007/s10529-005-7184-6ISI: 000231164000007PubMedID: 16091887Scopus ID: 2-s2.0-23844548756OAI: oai:DiVA.org:mdh-2154DiVA: diva2:114817
Available from: 2006-03-09 Created: 2006-03-09 Last updated: 2015-07-07Bibliographically approved

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