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|Title: ||Studies related to antibody fragment (Fab) production in Escherichia coli W3110 fed-batch fermentation processes using multiparameter flow cytometry|
|Authors: ||Want, Andrew|
Thomas, Owen R.T.
Hewitt, Christopher J.
|Keywords: ||Propidium iodide|
Bis-(1, 3-dibutylbarbituric acid) trimethine oxonol
|Issue Date: ||2009|
|Publisher: ||Wiley / © International Society for Advancement of Cytometry|
|Citation: ||WANT, A. ... et al, 2009. Studies related to antibody fragment (Fab) production in Escherichia coli W3110 fed-batch fermentation processes using multiparameter flow cytometry. Cytometry Part A, 75 (2), pp.148-154.|
|Abstract: ||Background: Microbiology is important to industry therefore rapid and statistically
representative measurements of cell physiological state, proliferation and viability are
essential if informed decisions about fermentation bioprocess optimisation or control
are to be made, since process performance will depend largely upon the number of
metabolically active viable cells.
Methods: Samples of recombinant Escherichia coli W3110, containing the gene for
the D1.3 anti-lysozyme Fab fragment under the control of the lac based expression
system, were taken at various stages from fed-batch fermentation processes and
stained with a mixture of bis-(1, 3-dibutylbarbituric acid) trimethine oxonol and
propidium iodide (PI/BOX). Where appropriate, measurements of dissolved oxygen
tension (DOT), OD600nm and Fab concentration were made.
Results: Depending on time of induction the maximum amount of Fab accumulating
in the supernatant varied quite markedly from 1 – 4 μgml-1 as did subsequent cell
physiological state with respect to PI/BOX staining with a concomitant drop in
maximum biomass concentration.
Conclusion: Depending on point of induction a 4 fold increase in Fab production
could be achieved accompanied by a ~50% drop in maximum biomass concentration
but with a higher proportion of viable cells as measured by multi-parameter flow
|Description: ||This is a pre-publication version of an article accepted for the journal, Cytometry Part A, and the definitive version is available from: http://www3.interscience.wiley.com/journal/121645129/issue|
|Version: ||Accepted for publication|
|Appears in Collections:||Published Articles (Chemical Engineering)|
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