Enhanced plasmid stability and production of hEGF by immobilized recombinant E. coli JM101

Chen, Xin-Ai; Xu, Zhinan; Chen, Peilin; Wong, Wan Keung

doi:10.1016/j.bej.2005.03.005

Cited by 26 publications

(17 citation statements)

References 13 publications

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“…In the immobilized cell system, the increase in productivity appeared to have resulted from both increased plasmid stability and increased specific productivity during continuous culture due to reduction in the probability of plasmid loss as a result of reduced specific growth rate, and perhaps a preferential retention of the plasmid-carrying cells in the fibrous-bed immobilized cell system. Similar increases in both volumetric and specific productivity have been demonstrated for protein production in immobilized E. coli JM101 cells (Chen et al, 2006).…”

Section: Discussionmentioning

confidence: 58%

“…Furthermore, by utilizing yeast proteins secretion systems, foreign peptides can be secreted to the external medium. In this respect, by utilizing yeast secretion systems, such as ␣-factor signal peptide, attempts were made to produce several recombinant proteins (Kilonzo et al, 2008;Gorgens et al, 2001;Bannister and Wittrup, 2000;Shiba et al, 1994;Vainio et al, 1994;Chau et al, 2000;Chen et al, 2006;Ekino et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

“…The application of the immobilization technique to bioprocess is widely accepted to have many advantages compared to conventional chemostat processes (Kilonzo et al, 2007a;Yang and Shu, 1996) and has been employed to minimize plasmid instability and to improve bioreactor productivity (Chau et al, 2000;Chen et al, 2006;Huang et al, 1996;Shu and Yang, 1996a;Yang and Shu, 1996;Barbotin, 1994;D'Angio et al, 1994). In the immobilized cell system, the maintenance of a plasmid-bearing cell population becomes easier as segregational instability and overgrowth of plasmid-free cells are less likely to happen because of reduced cell growth rate and selection between plasmid-carrying and plasmid-free-cell populations (Chau et al, 2000;Yang and Shu, 1996;Zhang et al, 1997a) and the compartmental cell distribution and mass transfer limitations in the immobilized cell environment .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Airlift-driven fibrous-bed bioreactor for continuous production of glucoamylase using immobilized recombinant yeast cells

Kilonzo

Margaritis

Bergougnou

2009

Journal of Biotechnology

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Section: Discussionmentioning

confidence: 58%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Airlift-driven fibrous-bed bioreactor for continuous production of glucoamylase using immobilized recombinant yeast cells

Kilonzo

Margaritis

Bergougnou

2009

Journal of Biotechnology

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“…EGF has an isoelectric point of pH 4.6 (Razis et al 2006) and molecular weight of 6045 Da (Cohen and Carpenter, 1975). Radioimmunoaassay (RIA) and enzyme-linked immunosorbent assay (ELISA) are the widely used methods for the quantification of EGF (Yamagata et al 1989;Kim et al 1992;Sivakesava et al 1999;Wang et al 2005;Chen et al 2006;Razis et al 2006).…”

Section: Introductionmentioning

confidence: 98%

Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance

Rosti¹,

Ramanan²,

Ling³

et al. 2013

Electron. J. Biotechnol.

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Background: A method for the selection of suitable molecular recognition element (MRE) for the quantification of human epidermal growth factor (hEGF) using surface plasmon resonance (SPR) is presented. Two types of hEGF antibody, monoclonal and polyclonal, were immobilized on the surface of chip and validated for its characteristics and performance in the quantification of hEGF. Validation of this analytical procedure was to demonstrate the stability and suitability of antibody for the quantification of target protein.Results: Specificity, accuracy and precision for all samples were within acceptable limit for both antibodies. The affinity and kinetic constant of antibodies-hEGF binding were evaluated using a 1:1 Langmuir interaction model. The model fitted well to all binding responses simultaneously. Polyclonal antibody (pAb) has better affinity (KD = 7.39e -10 M) than monoclonal antibody (mAb) (KD = 9.54e -9 M). Further evaluation of kinetic constant demonstrated that pAb has faster reaction rate during sample injection, slower dissociation rate during buffer injection and higher level of saturation state than mAb. Besides, pAb has longer shelf life and greater number of cycle run. Conclusions: Thus, pAb was more suitable to be used as a stable MRE for further quantification works from the consideration of kinetic, binding rate and shelf life assessment.

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“…Cell immobilization is now a widely accepted technique to minimize plasmid instability and to improve bioreactor productivity [1,4]. Over-production of a recombinant protein may unbalance metabolic processes and ultimately result in cell death because of the over-expression of a cloned gene or to the expression of a heterologous protein inside a bioreactor [1].…”

Section: Introductionmentioning

confidence: 99%

Repeated-batch production of glucoamylase using recombinant Saccharomyces cerevisiae immobilized in a fibrous bed bioreactor

Kilonzo

Margaritis

Bergougnou

2010

J Ind Microbiol Biotechnol

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The recombinant Saccharomyces cerevisiae strain C468/pGAC9 has an unstable hybrid plasmid pGAC9, which directs production of glucoamylase. A fibrous cotton material with a good adsorption capability for recombinant S. cerevisiae cells was used as the immobilization matrix in an internal loop airlift-driven fibrous bed bioreactor (ILALFBB) system. With batch cultures in the ILALFBB, the fraction of plasmid-carrying cells was 72% after more than 2 days cultivation, which was two times higher than that in the conventional free-cell culture. Correspondingly, a high activity of glucoamylase (GA; 113 U/l) was achieved with a high productivity of 43 U/l/h. The ILALFBB system also maintained a high fraction of viable plasmid-carrying of 74% for glucoamylase production during repeated-batch cultures, achieving a high glucoamylase activity of 140 U/l with a productivity of 19-130 U/l/h in all 14 batches studied during 19.8 days. The stable and long-term glucoamylase production from the ILALFBB was attributed to the effect of cell immobilization on plasmid stability. Plasmid-carrying cells were preferentially retained in the fibrous matrix because of their ability to adhere to the fiber surface and to form cell aggregates higher than those of plasmid-free cells. The repeated batch using immobilized cell of recombinant S. cerevisiae in the ALALFBB system thus provides a feasible method for stable, long-term and high-level production of glucoamylase.

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Enhanced plasmid stability and production of hEGF by immobilized recombinant E. coli JM101

Cited by 26 publications

References 13 publications

Airlift-driven fibrous-bed bioreactor for continuous production of glucoamylase using immobilized recombinant yeast cells

Airlift-driven fibrous-bed bioreactor for continuous production of glucoamylase using immobilized recombinant yeast cells

Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance

Repeated-batch production of glucoamylase using recombinant Saccharomyces cerevisiae immobilized in a fibrous bed bioreactor

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