• Title/Summary/Keyword: Immunoassay

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Molecular Cloning of H-Y Antigen Gene I. Purification of H-Y Antigen by Immunoaffinity Chromatography and Chemiluminescence Immunoassay for the Assay of H-Y Antigen (H-Y 항원 유전자의 cloning에 관한 연구 I. 친화성 크로마토그래피에 의한 H-Y 항원의 분리 정제 및 H-Y 항원 정량을 위한 화학발광 면역 분석법)

  • 김종배;김재홍;백정미;김창규;정길생
    • Korean Journal of Animal Reproduction
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    • v.15 no.2
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    • pp.149-155
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    • 1991
  • 본 실험은 H-Y 항원 유전자 크로닝을 위한 기초연구로서 H-Y 항원의 특성을 규명하기 위하여 친화성 크로마토그래피에 의하여 H-Y 항원을 분리·정제하였다. 정소 추출액을 항체가 결합된 column에 결합시킨 뒤 10% acetic acid로 용출시켰다. 용출된 분획을 모아 농축한 후 HPLC와 SDS-PAGE를 실시하여 H-Y 항원의 분자량은 약 6,7000달톤 임을 알 수 있었으며 isoelectric focusing에 의하여 등전점(pI)은 5.0인 것으로 측정되었다. H-Y 항원에 대한 단일클론항체와 표지항원으로는 H-Y 항원-ABEI(aminobutylethyl isoluminol)를 사용하여 H-Y 항원 정량을 위한 화학발광면역분석법을 개발하였다. 항원항체 반응후 빛의 측정은 NaOH 존재하에서 microperoxidase/H2O2를 이용한 산화반응으로 실시하여 10초간 측정한 빛의 양을 적분하였다. H-Y 항원의 농도와 빛의 양과는 역비례하였으며 감도는 11.8ng/tube 정도이었다.

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Time-resolved Fluoroimmunoassay for the Measurement of 17$\beta$-Estradiol using Anti-idiotypic Antibody (Anti-idiotype 항체를 이용한 17$\beta$-Estradiol 측정을 위한 Time-resolved Fluoroimmunoassay)

  • 김윤규;김창규;박성민;이치호;이원창;최영숙;김종배
    • Korean Journal of Animal Reproduction
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    • v.16 no.4
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    • pp.325-333
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    • 1993
  • A competitive type immunoassay method for 17$\beta$-estradiol(E2) based on the idiotypic anti-idiotypic antibody and time-resolved fluorescence is described. The anti-idiotypic antibody(Ab2) produced to E2 binding site of the primary idiotype antibody (Ab1) was labelled with europium and was allowed to compete with E2 standards or serum sample for the binding sites of Ab1 which was bound to 2nd antibody captured ontothe surface of microtitre plates. Fluorescence measured by time-resolved fluorometer was inversely proportional to the concentration of E2 over the range 5~500pg/well. The sensitivity of the assay was 5pg per well which was compatible with that ofradioimmunoassay using the same Ab1 and 3H-E2 as a tracer. One great advantage of this method described here was to enable antibodies to be labelled instead of haptens, and thus makes it easier to develop sensitive and robust immunoassay systems specially for haptens.

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Enzyme Immunoassay for Human Chorionic Gonadotropin Using Monoclonal Antibodies (단일크론성 항체를 이용한 융모막 성선자극 호르몬의 효소 면역측정법)

  • 차상훈;김희주;김원배;양중익
    • YAKHAK HOEJI
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    • v.31 no.2
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    • pp.64-69
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    • 1987
  • Monoclonal antibodies against human chorionic gonadotropin (hCG) were prepared and characterized by examining isotype, epitope binding, cross reactivity and affinity constants. And a sandwich type enzyme immunoassay for native hCG was developed with solid phase monoclonal antibody against the conformational determinant expressed only on native hCG and horseradish peroxidase conjugated monoclonal antibody against the $\beta$-subunit of hCG. The assay was sensitive to 1 mIU hCG/ml and shown a linear response up to 200 mIU hCG/ml. The cross reactivity for luteinizing hormone and $\beta$-subunit of hCG were 1% and 0.18%, respectively.

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Analytical Techniques Using ICP-MS for Clinical and Biological Analysis

  • Ko, Jungaa;Lim, H. B.
    • Mass Spectrometry Letters
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    • v.6 no.4
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    • pp.85-90
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    • 2015
  • This article reviews recent analytical techniques using inductively coupled plasma-mass spectrometry (ICP-MS) immunoassay for clinical and bio analysis. We classified the techniques into two categories, direct and indirect analysis, which depend upon a guideline of whether tagging materials are used or not. Direct analysis is well known, and generally used in conjunction with various other techniques, such as laser ablation, chromatographic separations, etc. Recently, indirect analysis using tagging elements has intensively been discussed because of its importance in future applications to bio and clinical analysis, including environmental and food industries. The method has shown advantages of multiplex detection, excellent sensitivity, and short analysis time owing to signal amplification and magnetic separation. Now, it expands the application field from small biomolecules to large cells.

SERS Immunoassay Using Microcontact Printing for Application of Sensitive Biosensors

  • Hong, Won-Jin;Seo, Hyeong-Kuyn;Jung, Young-Mee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4281-4285
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    • 2011
  • We introduced a promising patterned substrate by using a microcontact printing method that can be used for SERS immunoassays based on antigen-antibody binding. SERS spectrum of the Raman reporter with antibody, which is rhodamine 6G (R6G) adsorbed on colloidal gold nanoparticles, was observed only for the surfaces in which prostate-specific antigen (PSA) is present on the substrate that is attached to an immobilized layer of antibody on the gold nanoparticles layer of the patterned substrate. Raman mapping images clearly showed that the antibodies on the Raman reporter were successfully and selectively conjugated with the antigen on the patterned substrate. This method could be potentially extended to multi-protein detections and ultrasensitive biosensors.