Observation and modeling of induction effect on human transferrin production from stably transfected Drosophila S2 cell culture

Lim, Hye Jung; Joon, Hyung

doi:10.1016/j.enzmictec.2005.10.021

Cited by 22 publications

(24 citation statements)

References 27 publications

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“…In the last six years the Drosophila S2 protein expression system has been successfully used to produce a series of complex recombinant proteins (Kim et al 2008;Johansson et al 2007;Lim and Cha 2006;Cha et al 2005;Perret et al 2003;Cho et al 2004;Valle et al 2001). An analysis of the literature reveals that the major concern of researchers dealing with S2 cells has been the expression of high levels of the recombinant proteins, while there has little emphasis on collecting information about the influence of key factors such as cell metabolism or the composition of the medium on cell growth and biosynthesis of the recombinant protein.…”

Section: Introductionmentioning

confidence: 99%

Bioreactor culture of recombinant Drosophila melanogaster S2 cells: characterization of metabolic features related to cell growth and production of the rabies virus glycoprotein

et al. 2008

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Although several reports have been published on recombinant protein expression using Drosophila cells, information on their metabolism and growth in vitro is relatively scarce. In the present study, we have analyzed the growth and metabolism of transfected S2 cells (S2AcRVGP) in bioreactor cultures with serum-free medium Sf900 II, to evaluate its potential for mass production of a rabies virus glycoprotein (RVGP). Cells were cultured in a 3 lstirred-tank bioreactor at 28°C with pH controlled at 6.2 and dissolved oxygen at 50% air saturation. The cells attained a specific growth rate and maximum cell density as high as 0.084 h -1 and 2.3 9 10 7 cell ml -1 , respectively. The main substrates consumed during this rapid growth phase were glucose, glutamine and proline. An atypical accumulation of ammonia and alanine was observed in the culture medium, up to 62 mM and 47 mM, respectively, but lactate was produced in low levels. After exhaustion of glutamine and proline as energy sources, alanine was consumed and production of ammonia increased. The production of recombinant RVGP reached concentrations as high as 178 lg l -1 . Premature exhaustion of glutamine, serine and cysteine could be related to degradation of the recombinant glycoprotein. In general, the results demonstrated that S2AcRVGP can be considered an effective vehicle for large-scale recombinant glycoprotein expression and that several critical factors of the bioprocess could be optimized to increase the quality and productivity of the RVGP.

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Section: Introductionmentioning

confidence: 99%

Bioreactor culture of recombinant Drosophila melanogaster S2 cells: characterization of metabolic features related to cell growth and production of the rabies virus glycoprotein

et al. 2008

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“…It is currently recognized as one of the most promising systems for heterologous protein expression (Sorensen 2010). The Drosophila S2 protein expression system has been successfully used to produce numerous complex recombinant proteins (Moraes et al 2012;Cha et al 2005;Torfs et al 2000;Numao et al 2003;Nilsen and Castellino 1999;Chaiken et al 1995;Scotter et al 2006;Southern et al 1991;Culp et al 1991;Johansson et al 2007;Lim and Cha 2006;Perret et al 2003;Yokomizo et al 2007;Jorge et al 2008). In particular, the excellent yield obtained for the homologous srAFP, suggested the potential for strong expression of SSL (Scotter et al 2006).…”

Section: Discussionmentioning

confidence: 99%

Expression of recombinant Atlantic salmon serum C-type lectin in Drosophila melanogaster Schneider 2 cells

et al. 2012

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The Atlantic salmon (Salmo salar) serum lectin (SSL) is a soluble C-type lectin that binds bacteria, including salmon pathogens. This lectin is a cysteine-rich oligomeric protein. Consequently, a Drosophila melanogaster expression system was evaluated for use in expressing SSL. A cDNA encoding SSL was cloned into a vector designed to express it as a fusion protein with a hexahistidine tag, under the control of the Drosophila methallothionein promoter. The resulting construct was stably transfected into Drosophila S2 cells. After CdCl 2 induction, transfected S2 cells secreted recombinant SSL into the cell culture medium. A cell line derived from stably transformed polyclonal cell populations expressing SSL was used for large-scale expression of SSL. Recombinant SSL was purified from the culture medium using a two-step purification scheme involving affinity binding to yeast cells and metal-affinity chromatography. Although yields of SSL were very low, correct folding and functionality of the recombinant SSL purified in this manner was demonstrated by its ability to bind to Aeromonas salmonicida. Therefore, Drosophila S2 cells may be an ideal system for the production of SSL if yields can be increased.

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“…Many copies of recombinant plasmid vectors are inserted into the host cell genome, allowing continuous production of the desired foreign protein, after chemical induction, without cell destruction (Johanson et al, 1989;Kirkpatrick et al, 1995;Nilsen and Castellino, 1999;Shin and Cha, 2003;Lim and Cha, 2006). However, as with other insect cell lines, most heterologous glycoproteins produced by S2 cells show a simple paucimannosidic Abbreviations: Fuc, fucose; FucT, core ␣(1,6)-fucosyltransferase; Gal, galactose; GalT, ␤-1,4-galactosyltransferase; GalT, ␤(1,4)-galactosyltransferase; Glc, glucose; GlcNAc, N-acetylglucosamine; GlcNAcase, ␤-N-acetylglucosaminidase; GlcNAcT II, ␤(1,2)-N-acetylglucosaminyltransferase II; Man, mannose; Sia, sialic acid; SiaT, ␣(2,6)-sialylatransferase.…”

Section: Introductionmentioning

confidence: 99%

Expression of β-1,4-galactosyltransferase and suppression of β-N-acetylglucosaminidase to aid synthesis of complex N-glycans in insect Drosophila S2 cells

Kim

Kang

et al. 2011

Journal of Biotechnology

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Observation and modeling of induction effect on human transferrin production from stably transfected Drosophila S2 cell culture

Cited by 22 publications

References 27 publications

Bioreactor culture of recombinant Drosophila melanogaster S2 cells: characterization of metabolic features related to cell growth and production of the rabies virus glycoprotein

Bioreactor culture of recombinant Drosophila melanogaster S2 cells: characterization of metabolic features related to cell growth and production of the rabies virus glycoprotein

Expression of recombinant Atlantic salmon serum C-type lectin in Drosophila melanogaster Schneider 2 cells

Expression of β-1,4-galactosyltransferase and suppression of β-N-acetylglucosaminidase to aid synthesis of complex N-glycans in insect Drosophila S2 cells

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