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.
Wållberg
Fredrik
F
Department of Biotechnology, Royal Institute of Technology, Roslagstullsbacken 21, S-10691 Stockholm, Sweden.
Sundström
Helene
H
Ledung
Erika
E
Hewitt
Christopher J
CJ
Enfors
Sven-Olof
SO
D019149:Bioreactors / Q000382:microbiology; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry / Q000378:metabolism*; D007377:Interleukin-3; D008856:Microscopy, Fluorescence; D016185:Receptors, Interleukin-3 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D013926:Thrombopoietin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D005456:Fluorescent Dyes; D007377:Interleukin-3; D016185:Receptors, Interleukin-3; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C438891:promegapoietin-1a; D013926:Thrombopoietin