• Title/Summary/Keyword: 효소면역법

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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 an enzyme immunoassay for determination of steroid hormones to improve the reproductive efficiency of domestic animals (가축(家畜)의 번식효율증진(繁殖效率增進)을 위한 steroid hormones 의 효소면역분석법(酵素免疫分析法) 개발(開發))

  • Choi, Han-sun;Kang, Byong-kyu
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.611-615
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    • 1993
  • A rapid, solid-phase microtitre plate enzyme immunoassay(EIA) to determine the concentration of progesterone and testosterone in dairy cow is described. Both steroid hormones were analysed employing antibodies against $11{\alpha}$-hemisuccinate-progesterone bovine serum albumin and 4-androsten-$17{\beta}$-ol-3-one-carboxymethyloxime bovine serum albumin, respectively. as primary antibodies and sheep Ig G as secondary antibody. The conjugated used as labels for progesterone and testosterone was progesterone-$11{\alpha}$-hydroxy-hemisuccinate- horseradish peroxidase and 4 ${\alpha}$-androsten-$17{\beta}$-ol-3-hemisuccinate- horseradish peroxidase, respectively. Detection limit of microtitre plate EIA was 6.7 pg/well for progesteone and 1.0 pg/well for testosterone.

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Expression of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 in Radiation Exposed Small Intestinal Mucosa of the Rat (방사선조사를 받은 흰쥐 소장 점막의 손상과 재생과정 중 금속단백효소 및 억제자의 발현)

  • Kwag, Hyon-Joo;Lee, Kyoung-Ja;Rhee, Chung-Sik
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.66-74
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    • 2003
  • Purpose : The matrix metalloprotelnases (MMPs) are a family of enzymes whose main function is the degradation of the extracellular matrix. Several studies have revealed that MMPs and TIMPS are related to the wound heating process and in photoaging caused by ultraviolet Irradiation. However, the expressions of MMP and TIMP after irradiation have not, to the best of our knowledge, been studied. This study investigates the expressions of MMP-2 and TIMP-2 in rat Intestinal mucosa following irradiation. Materials and Methods : The entire abdomen of Sprague-Dawley rats was irradiated using a single dose method. The rats were sacrificed on day 1, 2, 3, 5, 7 and 14 following irradiation. Histopathological observations were made using hematoxilin & eosin staining. The expressions of MMP-2 and TIMP-2 were examined using immunohistochemistry, Irnrnunoblotting and ELISA. Results : Radiation induced damage associated with atrophic villi, and infiltration of inflammatory cell was observed from the first postirradiation day, and severe tissue damage was observed on the second and the third postirradiation days. An increase in mitosis and the number of regenerating crypts, as evidence of regeneration, were most noticeable on the fifth postirradiation day. From the immunohistochemlstry, the MMP-2 expression was observed from the first postirradiation day, but was most conspicuous on the third and the fifth postirradiation days. The TIMP-2 expression was most conspicuous on the fifth postirradiation day. From the irnrnunoblotting, the MMP-2 expression was strongly positive on the third postirradlatlon day, and that of TIMP-2 showed a strong positive response on the fifth postirradiation day. In ELISA tests, the expressions of MMP-2 and TIMP-2 were increased in the postirradiation groups compared to those of the normal controls, and showed a maximum increase on the fifth postirradiatlon day. These results were statistically significant. Conclusion : The expressions of MMP-2 and TIMP-2 were increased in the intestinal mucosa of the rats following irradiation, and these results correlated with the histopathological findings, such as tissue damage and regeneration. Therefore, this study suggests that MMP-2 and TIMP-2 play roles in the mechanisms of radiation-induced damage and regeneration of intestinal mucosa of rats.

Immunoelectrophoretic analysis of major component proteins In cystic fluid of Taenia solium metacestodes (면역전기영동법에 의한 유구낭미충 낭액의 구성 단백질 분석)

  • Yoon Kong;Seung-Yull Cho;Suk-Il Kim;Shin-Yong Kang
    • Parasites, Hosts and Diseases
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    • v.30 no.3
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    • pp.209-218
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    • 1992
  • When cystic fluid of Taenia solium metacestodes (CF) was filtrated through Sephacryl S-300 Superfine, major proteins were in fractions III add IV Major protein in fraction III was Band C protein of 150 kDa and that in fraction IV was Band N protein (Choi et of., 1990). When CF was electrophoresed in 0.9% agarose gel and reacted with anti-CF rabbit serum (RACF), two main bands, a long outer and a short inner band, were precipitated, together with 8 minor bands. RACF reacted with fraction III forming the long outer band whereas RACF formed the short infer band with fraction IV in immunoelectrophoresis (IEP) The long outer precipitin band of CF fraction III was similar to antigen B in hydatid fluid (HF) of Oriol et at. (1971), while the short inner band of CF fraction IV was similar to HF antigen 5 of Caption et at. (1967) . When HF was reacted with RACF, the short inner band was immunoprecipitated without forming the long outer band. Common antigenicity between CF and HF seemed to exist in fraction IV rather than in fraction III of CF. Patient sera of neurocysticercosis reacted more frequently with fraction III than with fraction IV.

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Serological Studies on the Specific Antibodies Against P-pili of Uropathogenic Escherichia coli (요로 감염환자에서 혈청학적 방법을 이용한 P-pili특이혈중 항체의 조사)

  • 이원용;김종배
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.31-40
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    • 1996
  • Escherichia coli is one of the most common etiological agents in urinary tract infection. An important virulence factor is the adhesive capacity of E. coli to uroepithelial cell, mediated by bacterial fimbriae. The Adhesion property has been regarded as an important virulence determinant in urinary tract infections. A total of 60 patients, who were diagnosed microbiologically as urinary tract infections, were examined by immunoblotting and enzyme-linked immunosorbent assay(ELISA). Uropathogenic E. coli with recombinant plasmid were positive for mannose resistant hemagglutination (MRHA). For identification of p-fimbriae subtype in uropathogenic E. coli, In the immunoblot analysis, specific bands in the range of p-fimbriae molecular weight of 17KD-22KD were identified. For the distribution of p-fimbriae subtype in the patient sera, 34/60(56.7%) were positive for $F7_1$, 28/60(46.7%) were positive for $F7_2$, and 30/60(50%) were positive for F13 with immunoblotting method. similar trends were observed in the enzyme-linked immunosorbent assay. Relatively good specificity(92.6%) and sensitivity(90%) were found in the ELISA test system using mixed antigens of purified $F7_1$, $F7_2$, and F13 p-fimbriae, and 60 sera from patients with urinary tract infections. In conclusion The serological tests were convenient method in diagnosis of urinary tract infections. among those ELISA could be recommended in diagnosis of urinary tract infections.

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시험관 내 면역 기술에 의한 항체 생산에 관한 연구

  • 전태훈;서동상
    • The Korean Journal of Zoology
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    • v.37 no.1
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    • pp.19-32
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    • 1994
  • 가장 효과적인 H-2b 항원에 대한 시험관내 항체 생산 조건을 찾기 위하여 근교계 생쥐인 C57BL/6BySnj의 비장세포를 UV로 불활성학 시킨 후 항원으로 사용하고, A/wySnJ$\times$Sm/J(ASmJF1, hybrid)의 비장세포를 항원 수용자 계통으로 하여 5-7일동안 배양기에서 항체 생산을 유도하였다. 본 실험에서 T 임파구 대식세포와 임파구 분화 촉진 인자인Concanavalin A Lipopolvsaccharide. Pokeweed mitogen 등을 사용하여 20가지 조건으로 실험을 수행하여, 항체 생성 여부는 보체 의존성 세포 장애 실험과 면역 효소법에 의해 조사하였다 그 결과 모든 조건에서 항체생산이 확인되었으며. 가장 좋은 시험관내 항체 생산 조건으로는 T 임파구와 대식세포를 함께 사용하여 면역시킨 것이 가장 효과적이었다. 이 방법을 이용하여 항체 생산을 유도한 후 5일째 면역된 비장세포를 Sp2/0-Ag 14와 세포 융합시켜 H-2b 마우스의 체포 표면 항원에 대한 단일군항체 생산을 시도하였다. 또한 생체내 면역 방법과 비교하기 위해 6주간 C57BL/6BySnJ의 비장세포를 복강내에 주사하여 같은 조건으로 세포융합을 시도하였다. 그 결과 H-2b 세포의 표면 항원에 대한 항체 생산을 하는 세포군은 시험관내 면역 방법에서 3개 생체내 면역 방법에서 4개부 확인되었다.

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Serological study on the cross-reactivity of bacteroides gingillalis, bacteroides intermedius and bacteroides asaccharolyticus by indirect immunofluorescence and enzym-linked immunsorbent assay (형광 현미경법 및 효소결합 면역협착법을 이용한 Bacteriodes gingivalis, Bacteriodes intermedius 및 Bacteriodes asaccharolyticus의 혈청학적 연구)

  • Chung, C.P.;Lee, J.Y.;Lee, Y.H.;Chung, H.W.;Chung, H.J.
    • Journal of Periodontal and Implant Science
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    • v.16 no.1
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    • pp.100.1-100.1
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    • 1986
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Detection of Viral Antigens in Stool Using EIA in Hospitalized Children and Clinical Implication (간접 효소 면역측정법을 이용한 입원 환아의 대변에서 바이러스 항원의 검출과 임상적 의의)

  • Min, Jung Hye;Seo, Jeong Wan;Park, Hye Kyung
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.7 no.2
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    • pp.143-152
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    • 2004
  • Purpose: The purpose of this study is to detect viral coproantigens in children who were hospitalized with acute diarrhea and to compare its association with clinical symptoms. Methods: Seventy-four stool samples were collected from children admitted to Ewha Mokdong Hospital from March 1996 to December 1999. The samples were frozen and analyzed for rotavirus, adenovirus, enterovirus, astrovirus, and calicivirus by enzyme immunoassay (EIA) with monoclonal antibody. 53 stool samples were collected from patients with diarrhea (diarrheal group) and 21 stool samples from patients hospitalized for reasons other than diarrhea (control group). Clinical features and laboratory findings were reviewed in both groups. Results: Among 74 stool samples, virus antigens were detected in 60 samples. Of the 60 virus-positive stool samples, 47 enterovirus, 26 rotavirus, 16 adenovirus, 11 astrovirus, and 11 calicivirus antigens were detected by EIA. Of the 60 virus-positive stool samples, 28 samples have one viral antigen, 30 samples have 2 or more viral antigens, and 2 samples showed a simultaneous infection of Salmonella group B and enterovirus. There was no relationship between the detected virus and clinical features. Conclusion: In this study, viral coproantigen and clinical symptoms were not associated. In the future, further larger scale studies are necessary.

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Study on the Anti-HLA Antibody Production Using in vitro Immunization Technique (시험관내 면역기술에 의한 항 HLA 항체 생산에 관한 연구)

  • 김혜원;서동상
    • The Korean Journal of Zoology
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    • v.38 no.2
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    • pp.186-195
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    • 1995
  • 사람의 항 HLA 단일군 항체 생산의 선결 조건인 가장 효과적인 시험관내 면역 조건을 확립하기 위해, 사람의 혈중 임파구를 함원으로 하여 마우스의 대식세포, 흉선세포. 이들의 조건배지, 그리고 임파구 촉진인자 등을 포함한 14가지의 다양한 배양 조건에서 세포를 배양하였으며, 마우스의 비장세포와 사람의 혈중 임파구에서 각각 항체 생산을 유도하였다. 항체의 생성 여부는 면역효소법(I섬SA)으로 조사하였다. 마우스의 비장세포는 모든 조건에서 다량의 항체가 검출되었으며, 사람의 혈중 임파구 분화에는 마우스의 조건배지보다 PWM. LPS와 같은 임파구 촉진인자가 효과적임을 알 수 있었으며. 특히 allogenic MLC(Mixed Lymphocyte Culture)에 의해 임파구 분화유도에 유용한 물질이 생성됨을 알 수 있었다.

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방역시스템 구축을 통한 살모넬라 청정화에 노력

  • 권준헌
    • KOREAN POULTRY JOURNAL
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    • v.37 no.4 s.426
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    • pp.72-75
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    • 2005
  • 1994년에 제정된 종계장위생관리요령과 1999년에 고시되었던 추백리 방역실시요령이 폐지되고 종계장$\cdot$부화장방역관리요령이 새롭게 만들어져 2004년 12월에 고시되었으며, 이중 검사시기, 검사방법 및 판정기준 등이 올 5월 1일부터 전면 시행될 예정이다. 이번에 제정 고시된 방역관리요령은 종계장$\cdot$부화장의 효율적인 방역관리와 추백리$\cdot$가금티푸스의 사전예방을 통해 양계농가에 건강한 병아리를 공급하기 위함이며, 이 고시에 수록된 주요 내용을 보면 추백리 및 가금티푸스에 감염된 종계군의 종계사용금지, 채혈시기 및 횟수, 종계혈청검사방법(급속전혈평판응집반응법, 효소면역법), 양성계군의 판단기준, 부화장에서 백세미용 알 및 병아리의 거래기록 작성 등이 새롭게 제시된 점이다. 종계장 및 부화장방역관리요령의 시행에 앞서 종계장 및 부화장에서의 살모넬라 감염증에 대한 피해를 최소화할 수 있는 방역관리에 대하여 알아보고자 한다.

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