• Title/Summary/Keyword: Hybridoma cell

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The Effect of Media Feeding Rate on the Production of Monoclonal Antibody Production in the Fed-batch Culture of Hybridoma (하이브리도마 세포의 유가식 배양에서 배지첨가속도가 단일클론 항체 생산에 미치는 영향)

  • 곽원재;최태부;박정극
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.272-280
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    • 1991
  • The effect of media feeding rate on cell growth and monoclonal antibody production in the fed-batch culture ot hybridoma A4W was studied. In the batch culture, the highest specific antibody production rate was observed at the begining of the culture period but its value tended to decrease rapidly with the culture time. The final antibody concentration and volumetric productivity was 65 $\mu g$/ml and 13 mg Mab/l/day, respectively. In the fed-batch culture, the specific antibody production rate, $q_p$ rebounded sharply within a few hours after the media feeding was started and it remained high until the end of culture if the media feeding was continued. The final antibody concentration was 220 $\mu g$/ml and the volumetric productivity was 45.1 mg/l/day. Further increase in final antibody concentration was achieved by applying a modified media of which component was fortified with glucose and glutamine, hence the final antibody concentration in this case was 270 $\mu g$/ml and the volumetric productivity was 51.8 mg/lday, which is as four tinlcs as high cuixparinf! to that of batch culture.

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Development of Direct Competitive Enzyme-Linked Immunosorbent Assay using Monoclonal Antibody (MAb) against Sulfamthazine (SMZ) and Establishment of Application Condition for Milk Sample (설파메타진에 단클론성 항체를 이용한 직접경쟁효소면역분석법의 개발과 우유 시료 적용 조건 확립)

  • Shim, Won-Bo;Mun, Chun-Sun;Kim, Jung-Sook;Choe, Ju-Mi;Kim, Ji-Hun;Park, Seon-Ja;Kang, Sung-Jo;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.176-182
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    • 2006
  • Sensitive and specific monoclonal antibody (MAb) was produced from hybridoma (1H11-5) obtained by fusion of myeloma cell (V653) and spleen cell isolated from mouse immunized sulfamthazine (SMZ)-HG-KLH. Direct competitive ELISA was developed for rapid detection of SMZ in milk samples using MAb against SMZ with optimized conditions between MAb and SMZ-HG-HRP conjugate, and applicable conditions for analysis of milk samples were established. Detection range of immunoassay was 0.1 to 100 ppb. Recoveries from spiked raw milk and processed milk samples averaged 82.1-120.7 and 82.1-97.1%, respectively.

Enhancement of Oxygen Transfer in Animal Cell Culture by Using a Perfluorocarbon as an Oxygen Carrier (과불소 탄소화물을 산소전달매체로서 사용하여 동물세포 배양의 산소전달 증진)

  • 조무환
    • KSBB Journal
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    • v.4 no.1
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    • pp.1-7
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    • 1989
  • In order to increase the oxygen transfer in a bioreactor for animal cell culture, a penluorocarbon, $Flutec^R$ ppll was used in a modified Celligen $Celligem^{TM}$. Also, the effects of ppll on the hybridoma cell growth and on the production of monoclonal antibody were investigated. There was no harmful effect of ppl1 on the cell growth and on the production of monoclonal antibody.

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Production of Monoclonal Antibody to Infectious Laryngo- tracheitis Virus by Cell Fusion (닭 전염성 후두기관염 바이러스에 대한 단크론성 항체생산)

  • Chung Ok Choi;Chung Gil Lee;Sung Man Cho;Soo Hwan An;Joon Hun Kwon
    • Korean Journal of Poultry Science
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    • v.15 no.3
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    • pp.199-206
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    • 1988
  • A total of 3 hybridoma clones producing monoclonal antibody (MCA) against infectious laryngotracheitis virus (ILTV) was established by somatic cell hybridization between mouse myeloma cells and spleen, cells from mice immunized with ILTV. The MCAs were screened by the indirect flourescent antibody (IFA) staining and the specific hybridomas were cloned by limiting dilution method. The MCAs produced by the 3 hybriomas were all classified as immunogloblin G and found to be reacting against common antigen(s) of high and low pathogenic ILTV examined. The titer of these antibodies in mouse ascitic fluid was from $10^5$ to $10^6$. Indirect fluorescent antibody test using these antibodies was found to be quite effective for the detection of ILTV from infected chickens being the most sensitive among the test methods adopted.

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Production of Monoclonal Antibody for Listeria spp. p60 Protein Based on iap Gene (Listeria spp. p60 단백질에 대한 단일클론항체의 생산)

  • 임희영;오연경;김종수;이영순;임윤규;윤병수
    • Journal of Food Hygiene and Safety
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    • v.18 no.1
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    • pp.25-29
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    • 2003
  • The p60 protein of Listeria spp. is a Listeria-Genus-specific, major extra-cellular protein, which is used as an indicator protein for the detection of these bacteria from contaminated foods. In this study, p60 protein were recombinantly produced in E. coli and were purified using amylose resin based column chromatography. Purified recombinant-p6O was used to generate monoclonal antibody against native p60. Antibody from hybridoma cell line, 1H4, specificically reacted with native p60 protein isolated from pathogenic Listeria spp. such as L. monocytogenes, L. ivanovii, L. welshimeri II, but did not or relatively weakly reacted with non-pathogenic Listeia species, L. innocua or other bacterial proteins. Antibody from 1H4 was produced using ascites fluid method and it may be useful to develop the Listeria-detection kits based on immunological method.

Characterization of Immunogens of Infectious Hematopoietic Necrosis Virus Isolated in Korea (전염성 조혈기 괴사 바이러스(IHNV)의 항원 유도 단백질 특성)

  • Park, Myoung-Ae;Sohn, Sang-Gyu;Park, Jeong-Woo;Jeong, Young-Kee
    • Journal of fish pathology
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    • v.7 no.1
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    • pp.13-22
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    • 1994
  • To identify the immunogens of a PRT strain of Infectious Hematopoietci Necrosis Virus (IHNV) isolated from cultrued fish in Korea (Park et al, 1993). a panel of 4 monoclonal antibodies (MAbs) against IHNV-PRT strain and two polyclonal antisera from rainbow trout survived IHN disease were prepared. Proteins of purified IHNV-PRT strain were analysed on 10% SDS-PAGE and transferred onto NC paper and were incubated with the antibody solutions. With the polyclonal antibodies, four bands ($M_1$, $M_2$, G and 90Kd) were detected and the band density was in the order of $M_2$ > 90Kd > $M_1$ > G. However, with the MAbs, only two bands(G and 90Kd) were detected. The origin of 90Kd protein was not clear but maybe cell. All the results represented that among the five proteins of IHNV-PRT strain (Park et al., 1993), $M_2$, $M_1$ and G proteins were immunogens and $M_2$ protein was the strongest one.

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Protective Effect of Extracts from Euryale ferox against Glutamate-induced Cytotoxicity in Neuronal Cells

  • Lee, Mi-Ra;Kim, Ji-Hyun;Son, Eun-Soon;Park, Hae-Ryong
    • Natural Product Sciences
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    • v.15 no.3
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    • pp.162-166
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    • 2009
  • Oxczaasssaidative stress plays an important role in neuronal cell death, which is associated with neurodegenerative conditions such as Alzheimer's and Parkinson's disease. This study evaluated the neuroprotective effect of Euryale ferox (EF) extracts against glutamate-induced cytotoxicity in hybridoma N18-RE-105 cells. Specifically, neuroprotective effects of methanol and ethanol extracts were evaluated by the MTT reduction assay. The ethanol extracts of EF displayed dose dependent protection against neuronal cell death induced by 20 mM of glutamate. Furthermore, the ethanol extracts of EF was sequentially fractionated with hexane, diethyl ether, ethyl acetate, and water layer according to degree of polarity. The hexane fractions exhibited neuroprotective effect against glutamate-stressed N18-RE-105 cells. Overall, results suggest that EF extracts can potentially be used as chemotherapeutic agents against neuronal diseases.

Production of Monoclonal Antibodies against Vibrio parahaemolyticus and Development of High Sensitive Immuno-Selective Filtration Method (Vibrio parahaemolyticus에 대한 단클론성 항체 개발과 고감도 면역선택여과법의 개발)

  • Kim, Jeong-Sook;Choi, Young-Dong;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.21-27
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    • 2016
  • The objectives of this study are to produce monoclonal antibodies (MAbs) against Vibrio parahaemolyticus and to develop an immuno-selective filtration (ISF) method for the rapid and sensitive detection of V. parahaemolyticus. The characterization of the MAb produced from HKVP 4H9-9 hybridoma cell was validated by enzyme-linked immunosorbent assay (ELISA) and western blot. The produced MAb was specific to V. parahaemolyticus and showed weak cross-reaction to V. alginolyticus, V. vulnificus and Staphylococcus aureus. After optimization of the method, $5{\times}10^1cell/mL$ of V. parahaemolyticus in a pure culture could be detectable. Although weak cross-reactivity to V. vulnificus, V. alginolyticus and Staphylococcus aureus was observed, the ISF was confirmed to be highly specific to V. parahaemolyticus. Especially, the ISF showed the most sensitivity compared to the immunoassays currently reported is easier to perform and quicker than ID-ELISA.

A Monoclonal Antibody That Specifically Binds Chitosan In Vitro and In Situ on Fungal Cell Walls

  • Schubert, Max;Agdour, Siham;Fischer, Rainer;Olbrich, Yvonne;Schinkel, Helga;Schillberg, Stefan
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1179-1184
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    • 2010
  • We report the generation of the first monoclonal antibody that specifically binds to the polysaccharide chitosan. Mice were immunized with a mixture of chitosans, and hybridoma clones were screened for specific binders, resulting in the isolation of a single clone secreting a chitosan-specific IgM, mAbG7. In ELISAs, the antibody could bind to chitosans of varying composition, but demonstrated the highest affinity for chitosans with lower degrees of acetylation (DA) and very poor binding to chitin. We tested the ability of the antibody to bind to chitosan in situ, using preparations of fungal cell walls. Immunofluorescence microscopy confirmed that the antibody bound strongly to the cell walls of fungi with high levels of chitosan, whereas poor staining was observed in those species with cell walls of predominantly chitin or cellulose. The potential use of this antibody for the detection of fungal contamination and the protection of plants against fungal pathogens is discussed.

Antibody Production in Plant Cell Cultures

  • Lee, James M.
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.67-78
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    • 1995
  • Monoclonal antibodies (MoAbs) are a highly diversified class of proteins with major research and commercial applications such as diagnostics and therapeutics. Currently, the dominant method for producing MoAbs is through the hybridoma technique. However, this technique is slow, tedious, labor intensive, and expensive. The production of MoAbs in cultured transgenic plant cells can offer some advantages over that in the over that in the mammalian systems. The media to cultivate plant cells are well defined and inexpensive. Contamination by bacteria or fungi is easily monitored in plant tissue cultures. Furthermore, these contaminants are usually not potent pathogens to human beings. In our interdisciplinary research efforts, heavy chain monoclonal antibody (HC MAb) was inserted into Ti plasmid vector and transferred into A. tumefaciens for the transformation in tobacco cells. It was found that 76% of the transformants produced HC MAb. The presence of HC MAb in the cell membrane fraction indicated that the signal peptide was functional and efficient. The change of the HC MAb concentration during a batch culture followed a similar trend as dry cell concentration, indicating that the production of HC MAb was growth related. The long-term repeated subcultures of 11 cell lines showed that there was no obvious trend of neither the decrease nor the increase of the productivity with the repeated subcultures.

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