High volumetric yields of functional dimeric miniantibodies in Escherichia
coli, using an optimized expression vector and high-cell-density
fermentation under non-limited growth conditions.
Functional bivalent miniantibodies, directed against the epidermal
growth factor receptor, accumulated to more than 3 gl-1 in high-cell-density
cultures of Escherichia coli RV308(pHKK) on a pilot scale. The miniantibodies
consist of scFv fragments with a C-terminal hinge followed by a
helix-turn-helix motif, which homodimerizes in vivo. The improved
expression vector pHKK is characterized by the hoklsok suicide system,
improving plasmid maintenance, and the inducible lac pl o promoter
system with the very strong T7g10 Shine-Dalgarno sequence. The expression
unit is flanked by terminators. The prototrophic RV308 cells were
cultivated in glucose mineral salt medium and reached a cell density
of 145 g dry biomass l-1 after 33 h. After induction, growth continued
almost unchanged for a further 4 h with concomitant miniantibody
formation. In the fedbatch phase, the concentration of glucose was
kept almost constant at the physiological level of approximately
1.5 gl-1, using on-line flow injection analysis for control. Surprisingly,
E. coli RV308(pHKK) did not accumulate significant amounts of the
metabolic by-product acetate under these unlimited aerobic growth
conditions.
%0 Journal Article
%1 citeulike:764851
%A Horn, U.
%A Strittmatter, W.
%A Krebber, A.
%A Knüpfer, U.
%A Kujau, M.
%A Wenderoth, R.
%A Müller, K.
%A Matzku, S.
%A Plückthun, A.
%A Riesenberg, D.
%C Hans-Kn��ll-Institut f��r Naturstoff-Forschung, Jena, Germany.
%D 1996
%J Appl Microbiol Biotechnol
%K expression ecoli
%N 5-6
%P 524--532
%T High volumetric yields of functional dimeric miniantibodies in Escherichia
coli, using an optimized expression vector and high-cell-density
fermentation under non-limited growth conditions.
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=9008885
%V 46
%X Functional bivalent miniantibodies, directed against the epidermal
growth factor receptor, accumulated to more than 3 gl-1 in high-cell-density
cultures of Escherichia coli RV308(pHKK) on a pilot scale. The miniantibodies
consist of scFv fragments with a C-terminal hinge followed by a
helix-turn-helix motif, which homodimerizes in vivo. The improved
expression vector pHKK is characterized by the hoklsok suicide system,
improving plasmid maintenance, and the inducible lac pl o promoter
system with the very strong T7g10 Shine-Dalgarno sequence. The expression
unit is flanked by terminators. The prototrophic RV308 cells were
cultivated in glucose mineral salt medium and reached a cell density
of 145 g dry biomass l-1 after 33 h. After induction, growth continued
almost unchanged for a further 4 h with concomitant miniantibody
formation. In the fedbatch phase, the concentration of glucose was
kept almost constant at the physiological level of approximately
1.5 gl-1, using on-line flow injection analysis for control. Surprisingly,
E. coli RV308(pHKK) did not accumulate significant amounts of the
metabolic by-product acetate under these unlimited aerobic growth
conditions.
@article{citeulike:764851,
abstract = {Functional bivalent miniantibodies, directed against the epidermal
growth factor receptor, accumulated to more than 3 gl-1 in high-cell-density
cultures of Escherichia coli RV308(pHKK) on a pilot scale. The miniantibodies
consist of scFv fragments with a C-terminal hinge followed by a
helix-turn-helix motif, which homodimerizes in vivo. The improved
expression vector pHKK is characterized by the hoklsok suicide system,
improving plasmid maintenance, and the inducible lac pl o promoter
system with the very strong T7g10 Shine-Dalgarno sequence. The expression
unit is flanked by terminators. The prototrophic RV308 cells were
cultivated in glucose mineral salt medium and reached a cell density
of 145 g dry biomass l-1 after 33 h. After induction, growth continued
almost unchanged for a further 4 h with concomitant miniantibody
formation. In the fedbatch phase, the concentration of glucose was
kept almost constant at the physiological level of approximately
1.5 gl-1, using on-line flow injection analysis for control. Surprisingly,
E. coli RV308(pHKK) did not accumulate significant amounts of the
metabolic by-product acetate under these unlimited aerobic growth
conditions.},
added-at = {2007-02-02T11:54:15.000+0100},
address = {Hans-Kn��ll-Institut f��r Naturstoff-Forschung, Jena, Germany.},
author = {Horn, U. and Strittmatter, W. and Krebber, A. and Kn{\"{u}}pfer, U. and Kujau, M. and Wenderoth, R. and M{\"{u}}ller, K. and Matzku, S. and Pl{\"{u}}ckthun, A. and Riesenberg, D.},
biburl = {https://www.bibsonomy.org/bibtex/227dafa99884baa6cbfa05dcda367f091/robert},
citeulike-article-id = {764851},
interhash = {bbad327d8fb60ef669ed1d93d62be52f},
intrahash = {27dafa99884baa6cbfa05dcda367f091},
issn = {0175-7598},
journal = {Appl Microbiol Biotechnol},
keywords = {expression ecoli},
month = {December},
number = {5-6},
pages = {524--532},
priority = {2},
timestamp = {2007-02-02T11:54:15.000+0100},
title = {High volumetric yields of functional dimeric miniantibodies in \textit{Escherichia
coli}, using an optimized expression vector and high-cell-density
fermentation under non-limited growth conditions.},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=9008885},
volume = 46,
year = 1996
}