Size characterization of inclusion bodies by sedimentation field-flow fractionation

Sedimentation field-flow fractionation (sedFFF) was evaluated to characterize the size of Δ(4–23)TEM-β-lactamase inclusion bodies (IBs) overexpressed in fed-batch cultivations of Escherichia coli. Heterologous Δ(4–23)TEM-β-lactamase protein formed different sizes of IBs, depending upon the induction...

Full description

Saved in:
Bibliographic Details
Published in:Journal of biotechnology 2008-11, Vol.138 (3), p.67-73
Main Authors: Margreiter, Gerd, Messner, Paul, Caldwell, Karin D., Bayer, Karl
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Chemistry
Escherichia coli
Escherichia coli K12
Fed-batch cultivation
Fractionation, Field Flow
Fundamental and applied biological sciences. Psychology
Inclusion Bodies
Kemi
NATURAL SCIENCES
NATURVETENSKAP
Particle Size
Reproducibility of Results
Sedimentation field-flow fractionation
β-Lactamase
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Sedimentation field-flow fractionation (sedFFF) was evaluated to characterize the size of Δ(4–23)TEM-β-lactamase inclusion bodies (IBs) overexpressed in fed-batch cultivations of Escherichia coli. Heterologous Δ(4–23)TEM-β-lactamase protein formed different sizes of IBs, depending upon the induction conditions. In the early phases of recombinant protein expression, induced with low concentrations of IPTG (isopropyl-β- d-thiogalactoside), IB masses were larger than expected and showed heterogeneous size distributions. During cultivation, IB sizes showed a Gaussian distribution and reached a broad range by the end of the fed-batch cultivations. The obtained result proved the aptitude of sedFFF to rapidly assess the size distribution of IBs in a culture.
ISSN:0168-1656
1873-4863
1873-4863
DOI:10.1016/j.jbiotec.2008.07.1995