• Title/Summary/Keyword: hybridoma cell line

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Ammonium Ion Effects and Its In Situ Removal by Using Immobilized Adsorbent in Hybridoma Cell Culture (하이브리도마 세포배양에서 암모늄 이온의 영향 및 고정화 흡착제에 의한 암모늄 이온의 동시제거)

  • 정연호;이해익
    • KSBB Journal
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    • v.11 no.3
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    • pp.329-339
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    • 1996
  • The effects of ammonium ion on cell growth kinetics, monoclonal antibody productivity, and cell metabolism of hybridoma cells were investigated. The mouse-mouse hybridoma cell line VlIIH-8 producing mouse IgG2a was used as a model system. Ammonium ion showed an inhibitory effect on cell growth and monoclonal antibody production. New immobilized adsorbents were developed for the reduction of the inhibitory effect of ammonium ion. The ammonium ion selective zeolite, Phillipsite-Gismondine was entrapped in calcium alginate bead or in dialysis membrane and applied to the hybridoma cell culture system for the in situ removal of ammonium ion from culture media. The effects of ammonium the both serum supplemented and serum free media on the cell growth were studied by applying immobilized adsorbents of calcium alginate bead type. The results demonstrated a substantial enhancement in cell growth. Applying immobilized adsorbents of dialysis membrane type to serum supplemented media also resulted in the stimulation of cell growth, cell viability and monoclonal antibody production.

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Monoclonal antibody의 대량 생산을 위한 hybridoma cell의 생존능 증가에 관한 연구

  • Ha, Seong-Jin;Im, Seon-Ha;Lee, Jong-Won;Jo, Mu-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.561-562
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    • 2003
  • Hybridoma cell is very important in point of producing monoclonal antibody(Mab). Producing large quantity of Mab is economically valuable. On this experiment, we used one of hybridoma cell line, 5F12 AD3, and treated various antibiotics such as genetitin(G418), ciprofloxacin and minocycline to improve cell viability and we expect that improving cell viability brings higher concentrations of Mab. The optimum concentration of each antibiotics for improving cell viability were 10ug/ml for G418, 1ug/ml or 10ug/ml for ciprofloxacin and 1ug/ml for minocycline.

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Fractionated Aged Black Garlic Extracts Enhance Growth of Anti-My-10 Hybridoma Cells and Production of IgG1 Antibody

  • Lee, Ji Young;Chung, Namhyun;Lee, Yong Kwon
    • Journal of Applied Biological Chemistry
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    • v.57 no.1
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    • pp.61-63
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    • 2014
  • Aged black garlic (ABG) was extracted with 20% ethanol and water (crude extracts) and fractionated into three categories (>10, 3-10, and <3 kDa). The effect of crude extract supplements on anti-My-10 hybridoma cell growth and IgG1 antibody production was investigated in suspension culture with a chemically defined protein-free medium. We observed that supplementation of ABG to the cell culture medium stimulated anti-My-10 hybridoma cell growth and production of IgG1 antibody, particularly with fractionated ABG of low molecular weight. The stimulation depended upon the concentration and the size of the fractionated ABG. We also found that the growth-promoting activity was not correlated with high antibody production. These results suggest that fractionated ABG is a novel and promising alternative as an animal cell culture supplement.

Develpment of Low-serum Medium(LSM) for Mouse-mouse Hybridoma Part II. Development of Low Serum Medium by Screening for Serum Replacement (Hybridoma 배양을 위한 저혈청 배지의 개발 제2부: 혈청 대체 물질 선정을 통한 저혈청 배지 제조)

  • 제훈성;최차용
    • KSBB Journal
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    • v.7 no.2
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    • pp.96-101
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    • 1992
  • A low serum medium(LSM) suitable for the growth of a self-constructed hybridoma cell line, KA112, was established by selecting ingredients to replace serum. Insulin and sodium pyruvate was important for the growth of cell line KA112. Various basal media were tested and DMEM gave the most favorable result. Low serum medium (LSM) developed in this work showed cell line stability in the culture for more than 6 months and exhibited cell growth equivalent to that carried out in medium supplemented with 7% FBS. LSM was found to be applicable to the suspension culture of KA112. The reduction of serum level down to 1%(V/V) FBS in LSM resulted in a substantial saving in the cost of media preparation for large scale culture.

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Low Serum Medium for CH07E02 Hybridoma (하이브리도마 배양을 위한 저혈청배지의 조성 결정)

  • 박상재;최차용
    • KSBB Journal
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    • v.8 no.5
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    • pp.473-477
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    • 1993
  • Up to now, 10% Fetal Bovine Serum(FBS(V/V)) was added to basal medium for the cultivation of hybridoma. For the cultivation of hybridoma cell line, CH07E02, against colon cancer, serum concentration was reduced to 3% FBS without influence on cell growth and maximum cell concentration. By the addition of cell growth promoting substances-insulin (I), pyruvate (P), oxaloacetate(O), Pluronic F-68(P) and 2-mercaptoethanol(2-ME)-to 1% FBS medium, a cell density higher than that with 1% FBS medium alone was achieved. FBS 3% medium was replaced by very cheap 2% Calf Serum (CS) medium without influence on cell growth rate and concentration. Cells grew vigorously in 0.5% CS+IPOP medium. This composition was used during suspension culture and exhibited good viability and high specific growth rate.

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Lipids Supply for Hybridoma Culture (하이브리도마 배양을 위한 lipids의 공급)

  • 박상재;최차용
    • KSBB Journal
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    • v.8 no.5
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    • pp.483-487
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    • 1993
  • BSA/acids component in serum free medium (SFM) developed for the culture of hybridoma cell line, KA112, was replaced by acids/Pluronic F-68 emulsion. Protein content of SFM was minimized, and increased maximum cell density was obtained in serum-free lipids supplemented medium (SFLSM). Cell growth promotion effect of the emulsion was not affected by filtration with 0.2$\mu$m filter.

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Development of Monoclonal Antibodies Specific to Galectin of Pine Wood Nematode, Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle and Their Utilization for Detection of Pine Wood Nematodes (소나무재선충[Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle]의 GaLectin에 대한 특이적인 단클론 항체 제작과 진단에의 활용)

  • Kim, A-Young;Kim, Young Ha;Choi, Bo-Hye;Nguyen, Trang;Yoon, Kyungjae Andrew;Lee, Si Hyeock;Han, Hye-Rim;Koh, Young Ho
    • Korean journal of applied entomology
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    • v.57 no.1
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    • pp.7-13
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    • 2018
  • Currently, there is no available tool that rapidly diagnoses pine wood nematode (PWN)-infected pine trees in the field. In this study, we synthesized and purified PWN Galectin, which might be an antigen specific to PWN, using the Baculovirus expression system. We used PWN Galectin as an antigen for generating 1,464 fusion hybridoma cell lines secreting monoclonal antibodies (Mabs). Among them, we selected 62 fusion hybridoma cell lines showing high reactivity to PWN Galectin. We further selected 12 fusion hybridoma cell lines showing high reactivity to the standard PWN-infected pine tree phosphate buffered saline (PBS) extract. Additionally, two fusion hybridoma cell lines showing no or extremely low reactivity were used as controls. The selected fusion hybridoma cell lines were subjected to limiting dilutions for selecting and establishing Mab-secreting cell lines showing higher reactivity to the standard PWN-infected pine tree extract than to the standard normal pine tree PBS extract. Moreover, the selected fusion hybridoma cell lines were further selected based on their higher reactivity to PWN protein extracts than to three non-pathogenic nematode protein extracts. The Mab-secreting cell lines established in this study could be used to develop rapid diagnostic tools that can be used in the field or in laboratories for detecting PWN-infected pine trees or PWN.

Glucose Effects on Cell Growth, Antibody Production, and Cell Metabolism of Hybridoma Cells (Hybridoma 세포의 세포성장, 항체생산 및 세포대사에 미치는 Glucose의 영향)

  • ;Shaw S.Wang
    • KSBB Journal
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    • v.10 no.3
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    • pp.323-334
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    • 1995
  • The effects of glucose on cell growth kinetics, monoclonal antibody productivity, and cell metabolism or hybridoma cells were investigated. The mouse-mouse hybridoma cell line VIII H-8 producing mouse IgG2a was used as a modal system. Glucose showed substrate inhibition type dependence on specific growth raie. The maximum cell density increased as initial glucose concentration increased up to 4 g/$\ell$. Glucose showed a strong influence on cell death kinetics, and an inverse relationship between specific death rate and glucose concentration was found. Cell viability and monoclonal antibody production increased as initial glucose concentration increased. The specific glucose consumption rate increased with glucose concentration, and cumulative specific lactate production rate increased with increasing initial glucose concentration. The overall kinetics of ammonium ion production was almost invariant with respect to initial glucose concentration, while the cumulative specific ammonium ion production rate was dependent on initial glucose concentration.

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Influence of Amino Acid and Vitamin Addition on the Growth and Metabolism of a Hybridoma in Batch Culture (하이브리도마의 회분식배양에서 아미노산과 비타민의 첨가에 따른 세포성장과 대사의 변화)

  • 이동섭;박홍우
    • KSBB Journal
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    • v.13 no.3
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    • pp.289-294
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    • 1998
  • The effects of various step-fortifications of the initial medium with amino acids, glucose, and vitamines on the growth and metabolism of a hybridoma cell line in batch cultures were quantified. Comparisons between the metabolic rates of the various cultivations were made for the exponential growth phase. Fortification of the basal medium resulted in higher cell densities through a prolonged growth phase, but the maximum specific growth rate was not affected. The uptake rate of glutamine increased with the addition of amino acids but did not change upon the addition of glucose or vitamines. The specific glucose consumption decreased slightly with the addition of amino acids but increased production of lactate and {{{{ { NH}`_{4 } ^{ +} }}}}. A reciprocal relationship between the yields of {{{{ { NH}`_{4 } ^{+ } }}}} and lactate indicated a joint regulation of glycolysis and glutaminolysis.

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Production of Monoclonal Antibody against Zearalenone Produced by Fusarium sp. (Fusarium sp.가 생성하는 zearalenone에 대한 단크론성 항체생산)

  • Kang, Sung-Jo;Chung, Duck-Hwa;Kang, Jin-Soon
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1409-1414
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    • 1998
  • To develop zearalenone-specific monoclonal antibodies, hybridoma cells were produced by fusion of myeloma cells $(P3{\times}63Ag\;V653)$ and spleen cells from BALB/c female mice immunized with zearalenone-oxime coupled to bovine serum albumin (BSA). After screening of antibody titer of them with a sandwich type enzyme-linked immunosorbent assay (ELISA), 5 hybridomas which could produced monoclonal antibodies with a high affinity for zearalenone were selected. The monoclonal antibody produced by Z-2-M26 hybridoma exhibited the high sensitivity to zearalenone and a little cross-reactivity to ${\alpha}-zearalenol$ (11%), but did not react with ${\beta}-zearalenol,\;{\alpha}-zearalenol,\;{\beta}-zearalenol$ and DON. In conclusion, the developed monoclonal antibody appeared to be a very promising immunoreagent for the future development of a specific and sensitive quantitative ELISA for zearalenone.

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