• Title/Summary/Keyword: cell production

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Continuous Stable production of won Willerand Factor Monoclonal Antibody in Spin Filter Bioreactor with Bleeding Technology

  • Yun, Joung-Won;Lee, Soo-Young;Park, Byung-Wook;Han-Kyu oh;Kim, Se-Ho;Byum, Tea-Ho;Park, Soung-yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.130-135
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    • 2000
  • The characteristics of two different modes of perfusion culture, intermittent and continuous bleedings, were investigated by culturing the hybridoma cells producing von Willebrand Factor (vWF) monoclonal antibody (McAb) in a 15 L bioreactor without clogging the filter. Both culture methods exhibited similar profiles of cell density and metabolite concentrations during the culture period at the cell concentration of around 1${\times}$107 cells/mL. When the perfusion rate was increased, the intermittrnt bleeding culture showed problems of ammonia accumulation and decrease of cell viability. The continuous bleeding culture in terms of nutrient consumption and metabolite production kinetics. But the analysis of specific oxygen consumption rate showed that the specific oxygen consumption rate of intermittent bleeding culture was similar to that of exponential growth phase. The continuous bleeding culture showed higher specific oxygen consumption rate of intermittent bleeding culture. finally we proved the possibility of long-term operation of continuous bleeding culture and produced approximately 40 g of vWF McAb in a 15L bioreactor after one-month operation.

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Optimal Conditions for the Production of Sphimin, a Sphingomyelinase Inhibitor from Steptomyces sp. F50970

  • Sipkyu Lim;Park, Wan
    • Journal of Life Science
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    • v.9 no.2
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    • pp.5-8
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    • 1999
  • We isolated a sphingonyelinase (SMase) inhibitor, which would be a potential reagent to regulate cell proliferation, oncogenesis, and inflammation, from a strain of Streptomyces sp.. In this paper, we report the optimal conditions for the production of SMase inhibitor, designed as sphinin, from Streptomyces sp. F50970. The optimal carbon and nitrogen source were 1% soluble starch and 0.05%-0.15% trypton. Most of monosaccharides and high concentration of soluble starch above 1.0% caused falling of pH and sphinin production. Zn2+, Cu2+, Fe2+, Mn2+, and Co2+inhibited cell growth and the production of sphinin. Inorganic phosphate promoted the sphinin production. Optimal initial pH for the production of sphinin was 7.5-8.0. Addition of CaCO3 to the medium resulted in an increase of inhibitor production. Based on these results, we designed a fermentation medium for the production of a SMase inhibitor, sphinin, from Streptomyces sp. F50970.

Effects of high Cell Density on growth-Associated Monoclonal Antibody Production by Hybridoma T0405 Cells Immobilized in Macroporous Cellulose carriers

  • Hideki Mochoda;Wang, Pi-Chao;Fr Jr. Nayve;Ryuji Sato;Minoru Harige;Nakao Nomura;Masatoshi Matsumura
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.110-117
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    • 2000
  • Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.

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The Influence of Microinjection of Foreign Gene into the Pronucleus of Fertilized Egg on the Preimplantation Development, Cell Number and Diameter of Rabbit Embryos

  • Makarevich, A.V.;Chrenek, P.;Fl’ak, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.2
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    • pp.171-175
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    • 2006
  • The aim of this in vitro study was to test the effect of microinjection (Mi) of foreign gene into the rabbit egg pronucleus and epidermal growth factor (EGF) addition on the blastocyst rate, the cell number and the diameter of embryos, and to determine possible relationships between embryo cell number and embryo diameter. Blastocyst rate was significantly decreased in gene- Mi (G-Mi/E0) group (63.1%) comparing to intact ones (83.5%, $p_1$<0.05). The addition of EGF at 20ng/ml (G-Mi/E20) or 200 ng/ml (GMi/ E200) to gene-Mi embryos did not affect blastocyst rate (65.6 and 55.2% resp.). As a control for Mi, the eggs were microinjected with the same volume of phosphate-buffered solution (PBS-Mi) instead of the gene construct solution. Cell numbers and embryo diameters were measured from embryo images obtained on confocal laser scanning microscope. Bonferroni-modified LSD test showed that the embryo cell number in PBS-Mi group was significantly lower ($p_1$<0.05) and in gene-Mi group was tended to decrease compared with intact embryos. Embryo diameter was not different among experimental groups. No effect of EGF given at any doses both on the cell number and embryo diameter was found. A positive correlation between cell number and embryo diameter was observed in all groups of embryos. Since embryo diameter was not changed under the influence of Mi or EGF addition in this study, this seems to be more conservative characteristics of the embryo morphology. These results suggest that the pronuclear microinjection compromises developmental potential of embryos, decreasing blastocyst rate and embryo cell number, whilst embryo diameter is not affected. No effects of EGF on studied parameters were confirmed. Declined quality of Mi-derived embryos is caused by the microinjection procedure itself, rather than by the gene construct used.

High Density Culture of KA112 Hybridoma and Effect of Glucose Concentration on MAb Productivity (하이브리도마의 고농도 배양과 포도당 농도가 MAb 생산성에 미치는 영향)

  • 박상재;최차용
    • KSBB Journal
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    • v.8 no.5
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    • pp.478-482
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    • 1993
  • Perfusion culture was conducted in Celligen perfusion culture system using a self-constructed hybridoma cell and low serum medium. The culture system employed hollow fiber to separate cells from the culture broth. Maximum cell density of $2.1\times10^7$ ce11s/m1, 10 times higher than in batch culture, could be achieved. Concentration of monoclonal antibody (MAb) was 4 times higher and production rate at maximum feed rate was 9 times higher than in batch culture. Glucose concentration was very important for the cell growth and MAb production. When glucose concentration was below 1g/l, i. e. 0.5~0.9g/l, specific MAb production rate decreased but cell concentration still increased. As the glucose concentration goes above 1g/l, specific MAb production rate increased and remained at maximum value at more than 1.5g glucose/l. The maximum value of the specific Mab production rate was similar to that of batch culture.

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Aqueous Extract of Schizandra chinensis Suppresses Dextran Sulfate Sodiuminduced Generation of IL-8 and ROS in the Colonic Epithelial Cell Line HT-29

  • Lee, Young-Mi;Lee, Kang-Soo;Kim, Dae-Ki
    • Natural Product Sciences
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    • v.15 no.4
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    • pp.185-191
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    • 2009
  • Intestinal epithelial cells (IEC) play an important role in the mucosal immune system. IEC-derived mediators of inflammatory cascades play a principal role in the development of colon inflammation. The aim of this study was to investigate the inhibitory effect of aqueous extracts of Schizandra chinensis fruits (SC-Ex) on the production of inflammatory mediators by the human colonic epithelial cells. HT-29 cells were stimulated with dextran sulfate sodium in the presence or absence of SC-Ex to examine the cytoprotection and production of IL-8 and reactive oxygen species (ROS). It was shown that dextran sulfate sodium (DSS) caused the reduction of cell viability and production of IL-8 and ROS in DSS-treated HT-29 cells. We observed that the treatment of SC-Ex protected significantly cell proliferation from DSS-induced damage in dose-dependent manner. SC-Ex (10 and 100 ${\mu}g$/ml) also suppressed DSS-induced production of IL-8 mRNA and protein. Moreover, DSS-induced ROS production was inhibited markedly by the treatment of 100 ${\mu}g$/ml SC-Ex. These results suggest that SC-Ex has the protective effects on DSS-induced cell damage and the release of inflammatory mediators in the intestinal epithelial cells.

Direct Synthesis of Dimethyl Ether From Syngas in Slurry Phase Reactor (액상 슬러리 반응기에서 합성가스로부터 DME 직접 제조)

  • Hwang, Gap-Jin;Kim, Jung-Min;Lee, Sang-Ho;Park, Chu-Sik;Kim, Young-Ho;Kim, Jong-Won
    • Journal of Hydrogen and New Energy
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    • v.15 no.2
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    • pp.119-128
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    • 2004
  • DME(Dimethyl Ether) was directly produced from the synthesis gas using the slurry phase reactor. The catalyst for DME production prepared two types (A type; Cu:Zn:Al=57:33:10, B type; Cu:Zn:Al=40:45:15, molar ratio). It was evaluated for the effect of the reaction medium oil using the small size slurry phase reactor. DME production yield and the methanol selectivity decreased in the order: n-hexadecane oil> mineral oil> therminol oil. The long-term test of DME production was carried out using A and B type catalyst, and n-hexadecane oil and mineral oil, respectively. It was confirmed that the use of A type for the catalyst and n-hexadecane for the reaction medium oil was very useful for the viewpoint of the DME production form the synthesis gas.

Production of Erythromycin Using a Carrier-Spported Mycelial Growth in a Fluidized-Bed Bioreactor (균사 증식 담체를 이용한 유동층 생물반응기에서 Erythromycin의 생산)

  • 김성환;배신규김정희
    • KSBB Journal
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    • v.4 no.3
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    • pp.241-245
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    • 1989
  • A carriersupported mycelial growth of Sreptomyces erythreus was applied to erythromycin fermentation sistem using celite as a support material. Hyphal growth through the pore matrices of the materials showed anchorages and provided a stable biofilm growth. When the phospate concentration was limited to 0.8g corn steep liquor/L(corresponding to 40mg KH2PO4/L), the specific production rate of erythromycin was increased from 557$\mu$g/g-cell.hr under unlimited condition to 2, 898 $\mu$g/g-cell.hr. A fluidized-bed bioreactor was operated for erythromycin production by a repeated fed-batch mode. The control of free mycelial concentration and the extension of production phase were considered important to maintain the reactor productivity at a desired level. The erythromycin production under phosphate-limited condition could be maintained for at least 600hrs.

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Effect of Inoculum Size on Biomass Accumulation and Ginsenoside Production by Large-Scale Cell Suspension Cultures of Panax ginseng

  • Thanh Nguyen Trung;Murthy Hosakatte Niranjana;Yu Kee-Won;Jeong Cheol Seung;Hahn Eun-Joo;Paek Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.6 no.4
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    • pp.265-268
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    • 2004
  • Cell growth and ginseng saponin production by large-scale suspension (bioreactor) cultures of Panax ginseng were investigated under various inoculum sizes. Cell growth was low at an inoculum size of 40 g FW/L, and the maximum cell growth was obtained with increasing inoculum size up to 100 g FW/L. The cell density of 333 g FW/L and 12.7 g DW/L was obtained at inoculum size of 100 g FW/L after 30 days of cultivation. Maximum saponin production of $4.40\;\cal{mg/g}$ DW was achieved at 60 g FW/L of inoculum size. Thus, inoculum size 60 g FW/L was suitable for optimum biomass accumulation as well as saponin production during bioreactor cultivation of ginseng suspension cells.

A Prediction of Chip Quality using OPTICS (Ordering Points to Identify the Clustering Structure)-based Feature Extraction at the Cell Level (셀 레벨에서의 OPTICS 기반 특질 추출을 이용한 칩 품질 예측)

  • Kim, Ki Hyun;Baek, Jun Geol
    • Journal of Korean Institute of Industrial Engineers
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    • v.40 no.3
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    • pp.257-266
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    • 2014
  • The semiconductor manufacturing industry is managed by a number of parameters from the FAB which is the initial step of production to package test which is the final step of production. Various methods for prediction for the quality and yield are required to reduce the production costs caused by a complicated manufacturing process. In order to increase the accuracy of quality prediction, we have to extract the significant features from the large amount of data. In this study, we propose the method for extracting feature from the cell level data of probe test process using OPTICS which is one of the density-based clustering to improve the prediction accuracy of the quality of the assembled chips that will be placed in a package test. Two features extracted by using OPTICS are used as input variables of quality prediction model because of having position information of the cell defect. The package test progress for chips classified to the correct quality grade by performing the improved prediction method is expected to bring the effect of reducing production costs.