Monitoring and quantification of inclusion body formation in Escherichia coli by multi-parameter flow cytometry

• 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.

Subject headings

NATURVETENSKAP  -- Biologi -- Biofysik (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences -- Biophysics (hsv//eng)

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Annan industriell bioteknik (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Other Industrial Biotechnology (hsv//eng)

Keyword

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

Biochemical process engineering

Publication and Content Type

ref (subject category)

art (subject category)