• Title/Summary/Keyword: hybridoma

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Characterization of Mouse Hybridoma Producing Monoclonal Antibody Paragonimus westermani (폐디스토마에 대한 Mmonoclonal Antibody생산 세포주개발에 대한 연구)

  • 고광삼;이숙영;이근배
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.64-68
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    • 1985
  • In this study, hybridoma techniques were applied to produce monoclonal antobodies to Paragonimus westermani, commonly known as lung distoma. Balb/c mice were immunized intraperitoneally every week with increasing doses of purfied protein of Paragonimus westermani adult worms begining with 0.3/mg/mouse/7 days and ending with 0.5mg at 28th day. Spleen cells from these immunized mice were hybridized with myeloma cells (NS-1) and the hybridized cells were selected in HAT media. The antibody secreting cells among the hybrid cells were initially selected by ELISA. Those initially selected cells were further screened by the criteria of antibody producing activity, and seneral cell lines among them were further tested with immunodiffusion method. One hybridoma clone produced IgM and another clone produced IgG1. The supernatant of the hybridoma clone producing IgM had titer 1:64 and the hybridoma clone producing IgG1 had titer 1:256 measured by immunofluorescence technique.

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Production of ${\alpha}2-Macroglobulin$ by a T Cell Hybridoma (T 세포 하이브리도마에 의한 ${\alpha}2-Macroglobulin$의 생산)

  • Lee, Chong-Kil;Han, Seong-Sun
    • YAKHAK HOEJI
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    • v.38 no.6
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    • pp.715-720
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    • 1994
  • ${\alpha}2-macroglobulin$ $({\alpha}2-M)$ has been shown to have a variety of activities. One of those activities is the suppression of immune response. Characterization of the immunosuppressive factor secreted by a T cell hybridoma showed that ${\alpha}2-M$ was produced and secreted from the T cell hybridoma. ${\alpha}2-M$ was produced abundantly from the T cell hybridoma when cultured as ascites. The isolation and identification of the ${\alpha}2-M$ were studied using affinity chromatography and N-terminal amino acid sequencing. The extended observations were that the ${\alpha}2-M$ produced by the T cell hybridoma suppresses mixed lymphocyte reaction.

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Development of Serum-free Media for the Culture of Mouse Hybridoma (II) ; Determination of the Role of Each Component and a Minimum Composition Media (쥐 하이브리도마 세포배양을 위한 무혈청 배지개발(II) -각 성분의 역할과 최소배지의 결정-)

  • 곽원재;조보연;최태부
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.489-493
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    • 1989
  • The role of each supplement in serum-free medium KM3 for the growth of hybridoma and the production of monoclonal antibody was investigated. Transferrin, ethanolamine and bovine serum albumin were shown to be indispensable for the growth of four kinds of hybridoma tested in this work, especially transferrin for Alps 25-3, and ethanolamine for A4W and KW hybridoma. The addition of $\beta$-mercaptoethanol to the culture medium of HCGK showed a good influence of both the cell growth and the production of monoclonal antibody. Upon the experimental results, we suggested a serum-free medium containing a minimum composition for the culture of hybridoma.

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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.

Prevalence of Listeria monocytogenes and Related Species in Minimally Processed Vegetables

  • Cho, Sun-Young;Park, Boo-Kil;Moon, Kwang-Deog;Oh, Deog-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.515-519
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    • 2004
  • Listeria spp. were isolated from a total of 402 naturally contaminated domestic ready-to-eat (RTE) vegetable samples by the conventional Food and Drug Administration protocol and confinned by API-Listeria kit. Also, the susceptibility to 12 antibiotics, polymerase chain reaction (PCR) assay for virulence gene of pathogenic Listeria monocytogenes isolates, and in vitro virulence assay using myeloma and hybridoma cells from murine and human sources were tested. Among the samples, 17 samples (4.2%) were found to be contaminated with Listeria species. Among the 17 strains of Listeria spp. isolates, only 2 strains (11.8%) of L. monocytogenes and 15 strains (88.2%) of L. innocua were identified. Antibiotic susceptibility test showed that the Listeria spp. isolates were very susceptible to the antibiotics tested, except for nalidixic acid. Among 17 strains of Listeria spp., PCR analysis showed that 2 strains of L. monocytogenes isolates proved to have a virulence hly gene, but none of L. innocua had the hly gene. Also, hybridoma Ped-2E9 cells assay showed that only L. monocytogenes isolates killed approximately 95-99% hybridoma cells after 6 h, but L. innocua isolates had about 0-5% lethal effect. These results indicate that PCR assay with hly primer or hybridoma Ped-2E9 cells assay could be used as a good monitoring tool or in vitro virulence test for L. monocytogenes.

Kinetic Analysis of the Effect of Cell Density on Hybridoma Cell Growth in Batch Culture

  • Lee, Eun-Yeol
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.117-120
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    • 2002
  • The effect of cell density on cell growth was investigated in a suspension batch culture of hybridoma cells. The specific growth rate was found to increase with increasing initial cell density and then to decrease with further increases in initial cell density. In order to quantitatively describe the dependence of specific growth rate on cell density, a kinetic model is proposed, which satisfactorily represents the experimental data.

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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