Abstract：
:For proteome research, antibodies against a growing number of antigens must be generated and characterized. The high throughput generation of antibody fragments, using in vitro selection, requires bacterial expression of antibody fragments. This created a need to establish an expression method to improve the parallel production of many antibody fragments. In this study, we describe the development of a high throughput bacterial production method for single chain fragment variables (scFvs) using shaking flasks or the LEX™ bioreactor. We compared the influence of a set of production parameters on Escherichia coli production of four different scFv. The results led to an optimized protocol for the parallel production of multiple antibody fragments.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Miethe S,Meyer T,Wöhl-Bruhn S,Frenzel A,Schirrmann T,Dübel S,Hust Mdoi
10.1016/j.jbiotec.2012.07.011subject
Has Abstractpub_date
2013-01-20 00:00:00pages
105-11issue
2eissn
0168-1656issn
1873-4863pii
S0168-1656(12)00406-3journal_volume
163pub_type
杂志文章abstract：:In this paper we report the development of a recombinant strain of the yeast Pichia pastoris, which secretes an anti-carcinoembryonic antigen single chain Fv (scFv) antibody fragment to the culture supernatant as a biologically active protein, at levels of 1.2 g l(-1). The yeast scFv was purified by IMAC, with a final...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(99)00183-2
更新日期：2000-01-21 00:00:00
abstract：:The production of the blue pigment indigoidine has been achieved in the entomopathogenic bacterium Photorhabdus luminescens by a promoter exchange and in Escherichia coli following heterologous expression of the biosynthesis gene indC. Moreover, genes involved in the regulation of this previously "silent" biosynthesis...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2011.10.002
更新日期：2012-01-01 00:00:00
abstract：:Plasmid instability of a recombinant Saccharomyces cerevisiae C468/pGAC9 (ATCC 20690) was examined during continuous culture in a nonselective medium in an airlift bioreactor. The recombinant strain contained a 2-micron based shuttle vector pGAC9 and expresses Aspergillus awatnori glucoamylase gene under the control o...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(97)00057-6
更新日期：1997-05-23 00:00:00
abstract：:Many recent epigenetic studies utilize the advantages of CRISPR/dCas9 based tools in linking certain epigenetic modification with gene expression regulation. Various multifactorial diseases often contain changed epigenetic signatures at many loci, so tools for simultaneously targeting different loci would significantl...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2019.05.306
更新日期：2019-08-10 00:00:00
abstract：:Streptomyces sp. strain CFMR 7, which naturally degrades rubber, was isolated from a rubber plantation. Whole genome sequencing and assembly resulted in 2 contigs with total genome size of 8.248 Mb. Two latex clearing protein (lcp) genes which are responsible for rubber degrading activities were identified. ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.09.007
更新日期：2015-11-20 00:00:00
abstract：:Bacillus velezensis G341 can suppress plant pathogens by producing antagonistic active compounds including bacillomycin D, fengycin, and (oxy) difficidin. The complete genome sequence of this bacterium was characterized by one circular chromosome of 4,009,746bp with 3953 open reading frames. The genome contained 36 ps...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.07.005
更新日期：2015-10-10 00:00:00
abstract：:Three matrices were used for immobilizing the cytochrome c: Sepharose CL-4B, Silasorb SPH amine and a laboratory-prepared new matrix based on crosslinked triazine (2,4,6-tris(aminoethylamine)-1,3,5-triazine) (TAT). Cytochrome c was immobilized on the matrices by several procedures and the amount of incorporated cytoch...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.01.014
更新日期：2005-05-25 00:00:00
abstract：:The poly(beta-hydroxybutyrate) (PHB) biosynthetic genes of Ralstonia eutropha that are organized in a single operon (phaCAB) have been cloned in Escherichia coli, where the expression of the genes in the wild-type pha operon from plasmid pTZ18U-PHB leads to the formation of 50-80% PHB/celldry mass when the cells are g...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(98)00127-8
更新日期：1998-10-27 00:00:00
abstract：:Xylanase T-6 is a thermostable alkaline-tolerant enzyme that is produced by Bacillus stearothermophilus T-6. Xylanase T-6 was found to bleach pulp effectively at pH 9 and 65 degrees C and was used successfully on an industrial-scale mill trial. To facilitate the future characterization of the protein via X-ray analysi...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(96)01604-5
更新日期：1996-11-15 00:00:00
abstract：:Medium composition and cultivation conditions were constructed for the optimum production of leupeptin by Streptomyces exfoliatus SMF13. The production of leupeptin was related to mycelial growth, being optimum in the cultivation with glucose-excess, phosphate-limited, and casamino acids medium. However, leupeptin-ina...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(95)00061-t
更新日期：1995-08-15 00:00:00
abstract：:Porcine circovirus Cap protein production by P. pastoris with strong AOX promoter suffered with the problems with traditional pure methanol induction: (1) inefficient methanol metabolism; (2) extensive oxygen supply load; (3) difficulty in stable DO control; (4) low protein titer. In this study, based on the differenc...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.02.018
更新日期：2014-05-10 00:00:00
abstract：:Genetically engineered human osteosarcoma cells containing developmental endothelial locus-1 (del-1) gene were studied for production of Del-1, a protein that has the properties of an extracellular matrix protein and can regulate vascular morphogenesis and remodeling. Del-1 has been studied as a potential anti-angioge...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/s0168-1656(02)00053-6
更新日期：2002-07-17 00:00:00
abstract：:The purification of human antibodies in an aqueous two-phase system (ATPS) composed of polyethylene glycol (PEG) 6000 and phosphate was optimised by surface response methodology. A central composite design was used to evaluate the influence of phosphate, PEG and NaCl concentration and of the pH on the purity and extra...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2007.04.002
更新日期：2007-10-31 00:00:00
abstract：:Bioprocesses such as production of organic acids or acid hydrolysis of bioresources during biofuel production often suffer limitations due to microbial sensitivity under acidic conditions. Approaches for improving the acid tolerance of these microbes have mainly focused on using metabolic engineering tools. Here, we t...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.01.015
更新日期：2014-03-10 00:00:00
abstract：:Enterokinase, a two-chain duodenal serine protease, activates trypsinogen by removing its N-terminal propeptide. Due to a clean cut after the non-primed site recognition sequence, the enterokinase light chain is frequently employed in biotechnology to separate N-terminal affinity tags from target proteins with authent...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2013.10.022
更新日期：2013-12-01 00:00:00
abstract：:The growth inhibition of a murine hybridoma mediated by ammonium chloride was shown to vary with the pH of the culture medium. Values for the initial media concentration causing 50% growth inhibition (IC50) ranged from 4 mM to 7.6 mM as the pH was reduced from 7.8 to 6.8. A significant negative correlation was observe...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(90)90053-e
更新日期：1990-07-01 00:00:00
abstract：:Dynamic miniaturized human multi-micro-organ bioreactor systems are envisaged as a possible solution for the embarrassing gap of predictive substance testing prior to human exposure. A rational approach was applied to simulate and design dynamic long-term cultures of the smallest possible functional human organ units,...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2010.02.001
更新日期：2010-07-01 00:00:00
abstract：:Different systems are being studied and used to prepare recombinant proteins for pharmaceutical use. The blood, and still more the milk, from transgenic animals appear a very attractive source of pharmaceuticals. The cells from these animals are expected to produce well-matured proteins in potentially huge amounts. Se...
journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/0168-1656(94)90062-0
更新日期：1994-05-31 00:00:00
abstract：:Escherichia coli phytase was covalently immobilized on NHS-activated Sepharose High Performance. The pH dependence of the phytase activity was not influenced by immobilization, whereas stability against heat treatment was enhanced as a consequence of immobilization. Compared to the free phytase the immobilized enzyme ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(96)01505-2
更新日期：1996-07-18 00:00:00
abstract：:Each of the twelve enzymes for glycolytic fermentation, eleven from Escherichia coli and one from Saccharomyces cerevisiae, have been over-expressed in E. coli and purified with His-tags. Simple assays have been developed for each enzyme and they have been assembled for fermentation of glucose to ethanol. Phosphorus-3...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2011.09.019
更新日期：2012-01-01 00:00:00
abstract：:The circadian clock schedules processes in microalgae cells at suitable times in the day/night cycle. To gain knowledge about these biological time schedules, Neochloris oleoabundans was grown under constant light conditions and nitrogen limitation. Under these constant conditions, the only variable was the circadian ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2014.07.016
更新日期：2014-10-10 00:00:00
abstract：:Mining sequence data is increasingly important for biocatalysis research. However, when relying on sequence data alone, prediction of the reaction catalyzed by a specific protein sequence is often elusive, and substrate specificity is far from trivial. The present study demonstrated an approach of combining sequence d...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2009.06.004
更新日期：2009-08-10 00:00:00
abstract：:Physiologically relevant in vitro models can serve as biological analytical platforms for testing novel treatments and drug delivery systems. We describe the first steps in the development of a 3D human brain tumour co-culture model that includes the interplay between normal and tumour tissue along with nutrient gradi...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2015.01.002
更新日期：2015-07-10 00:00:00
abstract：:Extracellular enzymes are assumed to be responsible for the initial and rate limiting step in biodegradation of polymers. Mainly enzymes with aliphatic esters as their natural substrates (e.g. lipase, cutinases) have until now been evaluated for polyester hydrolysis studies. However, the potential of enzymes with arom...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2017.01.012
更新日期：2017-09-10 00:00:00
abstract：:Clostridium thermocellum encodes a xylanase gene (xynC) which is the major component of its cellulosome. XynC is a multidomain enzyme comprising of a substrate binding domain at the N-terminal followed by the catalytic domain and a dockerin domain. To study the influence of binding domain on activity, stability and ex...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2010.07.021
更新日期：2010-10-01 00:00:00
abstract：:Izumoring is a new concept for the bioproduction of all hexose sugars - 16 aldohexoses, eight ketohexoses and 10 hexitols -- using enzymatic and microbiological reactions. The biocatalysts necessary for the strategy were (i) d-tagatose 3-epimerase [which epimerizes all ketohexoses at C-3 of the sugar], (ii) polyol deh...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2006.04.016
更新日期：2006-08-05 00:00:00
abstract：:Previously, we showed that the enzymes aspartokinase (AK) and dihydrodipicolinate synthase (DDPS), which are involved in L-lysine biosynthesis in the Gram-negative obligate methylotroph Methylophilus methylotrophus AS1, were inhibited by allosteric effectors, including L-lysine. To elucidate further the regulation of ...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2005.12.026
更新日期：2006-07-13 00:00:00
abstract：:A serum-dependent and two serum-independent variants of the Bowes melanoma cell line, RPMI7272, were transfected with plasmids containing a geneticin-resistance (neo) gene transcribed by the HSV thymidine kinase promoter and an SV40 T antigen gene under control of the mouse metallothionein I promoter. T-antigen increa...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(92)90014-z
更新日期：1992-11-01 00:00:00
abstract：:New antifungal compounds that circumvent the resistance of the pathogen by directly damaging yeast cell surface structures are promising agents for the treatment of fungal infections, due to their different mechanism of action from current clinically used antifungal drugs. We present here a rapid and cost-effective fl...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/j.jbiotec.2016.06.023
更新日期：2016-09-10 00:00:00
abstract：:We previously reported that a recombinant strain, SB412(pLGM304), was constructed from acidomycin-resistant mutants of Serratia marcescens and produced 200 mg of d-biotin per liter of a medium containing sucrose and urea (Sakurai et al., 1993a, b). In the present work, we intended to improve the d-biotin production. B...
journal_title：Journal of biotechnology
pub_type： 杂志文章
doi：10.1016/0168-1656(94)90024-8
更新日期：1994-07-29 00:00:00
