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Study of recombinant protein product...
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Ecole Polytechnique, Montreal (Canada).
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Study of recombinant protein production from transgenic Nicotiana tabacum plant cells in a perfusion bioreactor.
紀錄類型:
書目-語言資料,印刷品 : Monograph/item
正題名/作者:
Study of recombinant protein production from transgenic Nicotiana tabacum plant cells in a perfusion bioreactor./
作者:
Meneses Ramirez, Erick Alejandro.
面頁冊數:
49 p.
附註:
Source: Masters Abstracts International, Volume: 47-04, page: 2265.
Contained By:
Masters Abstracts International47-04.
標題:
Biology, Cell. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=MR46065
ISBN:
9780494460658
Study of recombinant protein production from transgenic Nicotiana tabacum plant cells in a perfusion bioreactor.
Meneses Ramirez, Erick Alejandro.
Study of recombinant protein production from transgenic Nicotiana tabacum plant cells in a perfusion bioreactor.
- 49 p.
Source: Masters Abstracts International, Volume: 47-04, page: 2265.
Thesis (M.Sc.A.)--Ecole Polytechnique, Montreal (Canada), 2008.
In a bioreactor culture, this cell line showed specific growth rates of 0.23 +/- 0.07/day in batch operation, and 0.29 +/- 0.11/day under perfusion operation (exponential growth period), reaching a maximum dry weight of 14.9 g I-1.
ISBN: 9780494460658Subjects--Topical Terms:
1017686
Biology, Cell.
Study of recombinant protein production from transgenic Nicotiana tabacum plant cells in a perfusion bioreactor.
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Thesis (M.Sc.A.)--Ecole Polytechnique, Montreal (Canada), 2008.
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In a bioreactor culture, this cell line showed specific growth rates of 0.23 +/- 0.07/day in batch operation, and 0.29 +/- 0.11/day under perfusion operation (exponential growth period), reaching a maximum dry weight of 14.9 g I-1.
520
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The use of whole plant cultures for the production of recombinant antibodies has recently received a great deal of attention due to many technical and economical advantages in comparison with microbial and mammalian production systems. These advantages include: inexpensive crop culture, easily scalable process, and low risk of contamination with mammalian pathogens.
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Plant cell culture of transgenic cell lines also offers a series of additional advantages over the whole plant approach, such as: precise control over feed nutrients and growth conditions, batch to batch consistency in product, and a high level of containment in the production of recombinant proteins. Despite of the previous advantages, production of recombinant antibodies in plant cell cultures exhibits a low yield compared to microbial and mammalian cell platforms. This is the main problem to overcome in order to produce recombinant proteins to be commercialized using a plant cell culture platform.
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In this master's project, a stable Nicotiana tabacum derived cell line (R514) was obtained. The objective of this project consisted in proving that the cell line R514 is able to produce and to secrete the murine immunoglobulin gamma 1 (IgG1) into the extracellular medium when cultured in a perfusion bioreactor. This recombinant protein was also recovered continuously from the extracellular medium using affinity chromatography.
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A feeding strategy that included nitrogen and phosphate limitation was also tried in a bioreactor culture. Nitrogen limitation was done so as to increase sugar accumulation. These carbohydrates serve as carbon skeletons for the synthesis of amino acids, required for the biosynthesis of proteins such as the immunoglobulin gamma 1 (IgG1). Phosphate limitation was intended to limit cellular growth. This limitation permits the increase of cultivation time, but also the limitation of cellular density in the bioreactor.
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The feeding strategy chosen permitted the production of about 3.49 mg of total IgG1 in a 3.0 litre bioreactor (2.7I of operation volume). Previous results are in contrast with 1.5 mg of total IgG1 obtained in a bioreactor conducted with full medium and without nutrient limitation. This leads us to believe that a posterior metabolic modulation of internal metabolite pools is an adequate alternative that deserves to be explored in order to increase the production of recombinant proteins in plant cell culture.
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