• Title/Summary/Keyword: 단클론 항체

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Preparation Monoclonal Antibodies against Bombyx Densonucleosis Virus Type II(Yamanashi isolate) (누에의 농핵병바이러스(산리주)에 대한 단클론항체의 제작)

  • 최홍규
    • Journal of Sericultural and Entomological Science
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    • v.32 no.1
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    • pp.44-48
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    • 1990
  • Monoclonal antibodies were prepared against Bombyx densonucleosis virus type-II(Yamanashi isolate). Four hybridoma clones, named C4, Fl, H2, M9 were only reacted with the DNV-II, but those were not reacted with Bombyx densonucleosis virus type-I(Ina isolate) and infectious flacherie virus(IFV) by double diffusion test in 0.8% agarose gel. C4, Fl and M9 of them were reacted with 53KDa polypeptide of DNV-II, and H2 was reacted with 46. 5KDa polypeptide of the virus.

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Establishment of B-1 cell-derived polyreactive monoclonal antibodies and expression of costimulators by B-cell to antigenic stimulation (B-1 세포 유래 다중반응성 단클론 항체의 형성과 항원 자극에 대한 B 세포의 동시자극자 발현)

  • Lee, Ju-Youn;Choi, Jeom-Il;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.371-384
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    • 2007
  • 연구 목적 : 면역 반응에서 B-1 세포의 정확한 역할은 아직 명확히 규명되지 않았으나 최근 B-1 세포가 면역의 내성을 야기하고 유지하는데 필요한 특성들을 가지고 있음이 밝혀지고 있다. 이에 본 연구에서는 B-1 세포 유래 단클론 항체의 특성과 항원 자극에 대한 B 세포의 동시자극자 (MHC Class, B7-2, 7-2) 발현을 평가하여 B 세포의 면역조절 기능을 알아보고자 한다. 연구 대상 및 방법 : B-1 세포 유래 단클론 항체를 형성하는 잡종세포주를 이용하여 다양한 내, 외인성 항원에 대한 단클론 항체의 반응 양상을 평가하였다. 여러 내, 외인성 항원으로 면역한 쥐의 복강과 비장 B 세포의 동시자극자의 발현을 Fluorescence Activated Cell Sorting (FACS)을 이용하여 평가하였다. 결과 : 최종적으로 단클론을 형성하는 2개의 클론을 형성하였고, 이 B-1 세포 유래 단클론 항체는 dose-saturable pattern을 띄는 다중 반응성을 나타내었다. FACS를 이용한 동시자극자의 발현 검사에서는 MHC 발현은 복강과 비장의 B 세포가 유사하였으나, B7-1과 7-2는 복강의 B 세포에서 더 뚜렷한 발현을 보여주었다. 결론 : B-1 세포 유래 단클론 항체는 다양한 내, 외인성 항원 자극에 대해 dose-saturable한 다중반응성을 나타낸다. 복강과 비장의 B세포는 내, 외인성 항원의 면역에 있어서 동시자극자 발현이 명확히 다른 양상을 나타냄을 확인할 수 있었다.

스쿠치카에 대한 단클론항체의 생물학적 특성

  • 정성주;오명주
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.517-518
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    • 2001
  • 스쿠치카는 넙치에 기생하는 섬모충으로 스쿠치카에 대한 숙주의 면역반응은 거의 알려져 있지 않다. 본 연구에서는 숙주의 스쿠치카에 대한 방어 메카니즘을 알기 위하여 스쿠치카에 대한 단클론항체를 만들어 스쿠치카에 대한 항체의 생물학적 특성을 보았다. (중략)

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Monoclonal antibody production for CP4 EPSPS detection assays (CP4 EPSPS 검출을 위한 단클론 항체 생산)

  • A-Mi Yoon;Il Ryong Kim;Wonkyun Choi
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.445-451
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    • 2021
  • In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

Passive transfer of immunity against Cryptosporidium infection in neonatal mice using monoclonal antibodies (단클론항체를 이용한 생쥐에서의 크립토스포리디움 감염의 수동면역)

  • 조명환
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.223-230
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    • 1993
  • Monoclonal antibodies (Ub) against merozoites and sporozoites of the protozoan parasite Cryptosporidium pcnjum were examined for potential modulation of cryptosporidial infections in vivo by daily oral Ub administration to oocyst-inoculated neonatal mice. Monoclonal-treated neonatal mice were sacrified four and eight days post infection (pi) . Differences in infection rates were observed among the treatment groups at the p < 0.05 level. Suckling mice treated daily with orally administered mixtures of mAbs( ascitic fluids) showed significantly reduced parasite loads compared to control mice at flour and eight days pi, while suckling mice receiving mAb Cmg-3 alone showed signiacant differences only at 4 days Pi., suggesting that passive transfer of mAb may be of value in controlling cryptosporidial infections.

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The Production and Characterization of a Monoclonal Antibody to Herpes simplex Virus Type 2 (Herpes simplex 2형 바이러스에 대한 단클론항체 생산과 항원 분석)

  • 최경은;이형환;조명환
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.97-104
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    • 1997
  • Herpes simpex virus type 2 (HSV-2) infects the genital and oral mucosae of human and other animals. HSV-2 infection is a widespread health problem causing various clinical syndromes including oral, genital, and ocular lesions, viral encephalitis, and recurrent diseases. Hybridorna cell lines secreting a monoclonal antibody (mAb) against the HSV-2 were produced by fusing spleen cells of HSV-2-immunized mice with Sp2/0-AgI4 myeloma cells. One hybridoma cell line was established and its monoclonal C-2, IgM, recognized the antigens of 134, 86, and 43 kDa in western blot analysis. In SDS-P AGE analysis of HSV -2 antigens, 25 bands were separated between 3D kDa and 159 kDa. In indirect immunofluorescent assay, mAbs exhibited binding to the virus antigen expressed on Vero cell infected with HSV-2.

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Immunological properties of the 30 kDa antigen of Toxoplasma gondii (단클론 항체를 이용하여 정제한 톡소포자충 30 kDa 항원의 면역학적 특성)

  • Lee, Yeong-Hwa;No, Tae-Jin;Sin, Dae-Hwan
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.55-62
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    • 1997
  • The molecular weight 30 kDa membrane protein of Toxoplusma Sondii (Toxoplasma 30 kDa) apparently conserved in most strains of T. gondii and sera of infected hosts. The present study aimed to elucidate Toxoplasmc 30 kDa as a useful diagnotic antigen for serodiagnisis of toxoplasmosis by ELISA and for induction of protective immunity. Murine spleen cells immunized with the membrane antigen of T. gondii were fused with mouse Sp2/0-Ag 14 myeloma cells. Out of 8 clones selected, five were IgG2b, the others belonged to IgG 1 and IgG2a. The 30 kDa antigen was distributed mainly on the surface membrane of tachyzoites by indirect fluorescence method. Murine peritoneal macrophages which were activated by 30 kDa antigen produced more amounts of NO2 compared with crude antigen-treated group, however there were no significant differences in toxoplamacidal activity between the two groups. Higher specificity of Toxoplosma 30 kDa antigen was recognized for serodiagnosis of toxoplasmosis than the crude antigen. From these results, ToxopLasmo 30 kDa antigen enhances the cytotoxic effect of macrophages as well as a more reliable means for the serodiagnosis of toxoplasmosis by ELISA. Key words: Toxoplosma gondii, 30 kDa antigen (p30), mouse, serodiagnosis, macrophage, cytotoxicity.

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Production of Monoclonal Antibody against Sulfamethazine and Development of ELISA

  • C.S. Chae;Lee, M.H.;Lee, H.;S.Y. Yoon;P.D. Ryu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.176-176
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    • 1996
  • 축산식품중에 잔류하고 있는 sulfamethazine을 검출하기 위하여 sulfamethazine에 대한 단클론항체를 생산하고 이를 이용하여 효소면역측정법을 개발하였다. 면역원은 sulfamethazine에 KLH를 그리고 흡착항원은 BSA를 glutaraldehyde법으로 결합시켰다. 면역원으로 Balb/c mouse를 면역시킨 다음 비장 형질세포률 얻어 myeloma cell과 융합하여 융합잡종세포를 만들었다. Sulfamethazine에 대한 항체를 분비하는 융합잡종세포를 단계회석법과 ELISA를 이용하여 cloning하여 D2, A9, B8, Bl 클론을 얻었다. 이들 클론에서 얻어진 단클론항체를 사용하여 indirect competitive ELISA를 실시하여 표준곡선을 작성하여 본 결과 농도의존성 곡선을 얻을 수 있었다. 4클론중에서 A9 클론을 사용하여 다른 유사한 sulfonamide듣과 p-aminobenzoic acid와 교차반응을 조사한 결과 sulfamerazine에 12.5%의 교차반응을 보였으나 다른 설파제에 대해서는 교차 반응을 보이지 않았다.

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Development of Enzyme-Linked Immunosorbent Assay for Glyphosate-Tolerant Soybeans (제초제내성 유전자재조합 콩의 검출을 위한 면역분석법 개발)

  • Kwak, Bo-Yeon;Ko, Seung-Hee;Park, Chun-Wuk;Son, Dae-Yeul;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.366-372
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    • 2003
  • Enzyme-linked immunosorbent assay (ELISA) for assaying the 5-enolpyruvyshikimate-3-phosphate synthase from Agribacterium sp. CP4 (CP4 EPSPS) in genetically modified soybeans was developed. Polyclonal and monoclonal antibodies (Pab, Mab) specific to the CP4 EPSPS were produced. When using the Pab, the detection limit of sandwich ELISA toward CP4 EPSPS (0.03 ${\mu}g/mL$) was better than that of competitive indirect ELISA(ciELISA) (1 ${\mu}g/mL$). It was found that 2 of 3 monoclonal antibodies, Mab1 and Mab2, recognized the same antigenic determinant on CP4 EPSPS, but Mab3 recognized different antigenic determinant when competitive ELISA was performed using the Mabs. On the other hand, when the sensitivity of sandwich ELISA using combination of Pab and/or Mabs was determined, the sandiwich ELISA using Mab2 as a capture antibody and Pab-HRP as a secondary antibody showed the lowest detection limit of CP4 EPSPS (0.02 ${\mu}g/mL$). The sandwich ELISA developed in this study could be applied to detect glyphosate-tolerant soybeans.

Monoclonal Antibody against leucocyte CD11b(MAb 1B6) increase the early mortality rate in Spraque Dawley with E. coli pneumonia (백혈구 CD11b에 대한 단 클론 항체 (MAb 1B6)는 Spraque Dawley의 E. coli 폐렴의 조기 사망률을 증가시킨다)

  • Kim, Hyung Jung;Kim, Sung Kyu;Lee, Won Young
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.4
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    • pp.579-589
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    • 1996
  • Background : Activation of neutrophil is critical for the clearance of microorganisms and toxic host mediators during sepsis. Unfortunately the activated neutrophil and its toxic byproducts can produce tissue injury and organ dysfunction. The leucocyte CD11/18 adhesion complex regulates neutrophil-endothelial cell adhesion, the first step in neutrophil migration to sites of injection and inflammation. To investigate the potential of neutrophil inhibition as a treatment strategy for sepsis, we evaluated the effects of monoclonal antibody against CD11b (MAb 1B6) in rats intrabronchial challenged with Escherichia coli. Methods : Animals were randomly assigned to receive monoclonal antibody against CD11b (1 mg/kg, sc) and bovine serum albumin(BSA, 1 mg/kg, sc) 6 hr before, at 0 and 6 hr after intrabronchial challenge of $20x10^9$ CFU/kg E. coli 0111. Animals were randomized to treat either 24, 60 or 90% oxygen after bacterial challenge and begining 4 hr after inoculation, all animals were received 100 mg/kg ceftriaxone qd for 3 days. Peripheral and alveolar neutrophil(by bronchoalveolar lavage) counts and lung injury parameters such as alveolar-arte rial $PO_2$ difference, wet to dry lung weight ratio and protein concentration of alveolar fluid were measured in survived rats at 12 hr and 96 hr. Results : Monoclonal antibody against CD11b decreased circulating and alveolar neutrophil especially more in 12 hr than in 96 hr The lung injury parameters of antibody-treated animals were not different from those of BSA-treated animals. but It was meaningless due to small number of survived animals. The early(6 hr) mortality rate was significantly increased in antibody-treated group(51%) compared to BSA-treated group(31%) (P=0.02) but late(from 12 hr to 72 hr) mortality rate was not different in antibody-treated group(44%) from BSA-treated group(36%) (P =0.089). Conclusion : Leucocyte CD11b/18 adhesion molecule is known to regulate neutrophil migration to the site of infection and inflammation. The monoclonal antibody against CD11b decreased alveolar neutrophil in rats with pulmonary sepsis and increased early mortality rate. Therefore, we can speculate that monoclonal antibody against CD11b blocks of alveolar recruitment of neutrophils, impairs host defense mechanism and increases early mortality rate of pulmonary sepsis in rat.

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