• 제목/요약/키워드: Chemical antigen

검색결과 125건 처리시간 0.025초

In Vitro에서 히스타민 측정 시 ELISA법과 HPLC-MS 분석법의 비교 (Comparison of Methods for Measuring Histamine by ELISA and HPLC-MS Assay In Vitro)

  • 이인희;김유현
    • 대한임상검사과학회지
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    • 제47권4호
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    • pp.306-312
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    • 2015
  • 알러지 반응이 일어나면 histamine이 방출되기 때문에 histamine을 정량 측정함으로써 유발된 알러지의 정도를 확인 할 수 있다. 일반적으로는 항원-항체반응으로 microplate reader를 이용하여 흡광도를 측정하여 정량 한다. 본 연구에서는 histamine 방출량을 측정함에 있어서 일반적인 항원-항체반응과 분석 화학적인 방법으로 HPLC-MS를 이용한 방법을 비교하였다. 세포주는 RBL-2H3를 사용하였고, C48/80으로 자극시켜 알러지를 유발하였다. 유발된 알러지는 ${\beta}$-hexosaminidase의 측정으로 탈 과립을 확인하였으며 실험의 정당성을 위하여 세포독성 능을 확인하였다. Histamine 정량에서 항원-항체반응에 의한 측정의 정량한계는 10.257 ppm이었고, HPLC-MS에 의한 정량한계는 0.020 ppm으로 현저한 차이를 보였다. 알러지 활성 및 항 알러지 실험에 있어서 histamine의 측정은 HPLC-MS를 이용한 분석이 더 정밀하고 정확한 실험인 것을 확인하였다.

Latex 응집반응을 이용한 동물의 톡소플라즈마병 진단용 kit 개발에 관한 연구 (Development of diagnostic kit(Test-MT) for the microplate latex agglutination test of toxoplasmosis in animal)

  • 서명득;주후돈;데이빗 마-스
    • 대한수의학회지
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    • 제35권3호
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    • pp.583-593
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    • 1995
  • The present study was conducted to develop a toxoplasma latex agglutination test antigen(Test-MT) and evaluate the toxoplasma latex agglutination(LA) test using a newly-made "Test-MT kit" by comparing with the Toxo-MT kit(Eiken chemical co, Tokyo). Also, the specifity and sensitivity test were made by comparing with IFA test and IgG-ELISA. Tachyzoite suspensions of Toxoplasma gondii(RH strain) were ultracentrifuged for 30min at $60,000{\times}g(4^{\circ}C)$ and the supernatant was used as a water-lysate antigen. Polystyrene latex particles of $1.0{\mu}m$ in diameter(Polyscience co) were used for the preparation of sensitized latex-antigen supension(Test-MT). The frequency distribution of LA titers in Test-MT showed two peaks at <1:32 and 1:128. The borderline titer for positive test in Test-MT was determined to be 1:64. But the frequency distribution of LA tites in Toxo-MT showed two peaks at <1:16 and 1:64. The positive borderline was determined to be 1:32. Agreement of reactions between Test-MT and Toxo-MT kit by LA test was shown 92.5% in bovine sera and 97.0% in swine sera, respectively. From the results obtained here it was determined that the sensitized latex-antigen, Test-MT kit, for the microtiter agglutination test prepared as same as by the procedure described in the previous paper(Suh and Lee, 1993) was useful as a highly specific, sensitive and stable immunotiteration reagent for serodiagnosis of toxoplasma infection in animal sera.

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Sprague-Dawley 랫트에 자연발생한 고환위축의 병리조직학적 관찰 (Histopathological observation of spontaneous testicular atrophy in Sprague-Dawley rat)

  • 손화영;김용범;하창수;강부현
    • 한국수의병리학회지
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    • 제2권1호
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    • pp.47-52
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    • 1998
  • The incidence of spontaneous testicular atrophy and its morphological changes during stage-specific spermatogenesis were investigated in male Sprague-Dawley rats at 10, 19, and 32 weeks of age. The incidence of testicular atrophy was 0.2%(2/90) 7.9%(9/114) and 10%(4/40) in 4, 13 and 26 weeks respectively. The epididymis with testicular atrophy had low sperm density. In the minimally affected tests scattered tubules showed complete depletion of germ cells without stage specificity. Testes with moderate to severe testicular atrophy showed seminiferous tubules lined with dense Sertoli cell population. While Leydig cells in the interstitium appeared hypertrophy they were immunohistochemically negative for proliferating cell nuclear antigen a marker of cell proliferation.

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피나스테라이드의 특성에 대한 이론적 연구 (Theoretical Study for the Characteristics of Finasteride)

  • 이철재;정맹준;김동엽
    • 한국산업융합학회 논문집
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    • 제16권3호
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    • pp.69-74
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    • 2013
  • Finasteride, ($5{\alpha}$, $17{\beta}$)-N-(1,1-dimethylethyl)-3-oxo-4-azaandrost-1-ene-17-carboxamide), is a 5a-reductase enzyme inhibitor. This enzyme converts testosterone to the more potent androgen, a-dihydrotestosterone. This molecules a logical medical treatment for benign prostatic hyperplasia (BPH), as it induced a reduction in serum dihydrotestosterone and prostatic specific antigen levels with a concomittant increase in blood testosterone concentration . Despite its widespread use, little has been published concerning its molecular properties. Therefore, in this study, in order to explain characteristics of finasteride, total energy, net charge, vibrational mode of melatonin are calculated by PM3 methods of HyperCam 8.0.

The Coordination of Pyridyl-N to Pentacyanoferrate for the Electrochemical Detecting Small Organic Molecules

  • Choi, Young-Bong;Jeon, Won-Yong;Kim, Hyug-Han
    • Bulletin of the Korean Chemical Society
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    • 제34권2호
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    • pp.595-599
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    • 2013
  • The coordination of pyridyl-N to pentacyanoferrate for the detection of small organic antigens in solution is presented. The unique contribution of this paper is the direct conjugation of pyridyl-N in small organic antigens to pentacyanoferrate. Pentacyanoferrate is promising as an electrochemical label owing to its good electro-chemical properties, which can be utilized to generate an electrical signal in homogeneous electrochemical immunoassays. The facilely synthesized pyridyl-N to pentacyanoferrate was characterized by the electrochemical and spectroscopic methods. Hippuric acid (HA) has been detected competitively on the interaction of free HA and pentacyanoferrate-(4-aminomethylpyridine-hippuric acid) (Fe-HA) to its antibody, with the detection limit of 0.50 ${\mu}g\;mL^{-1}$. While pentacyanoferrate-based immunoassay is in its simplicity and infancy, the proposed immunoassay offers attractive opportunities for developing pyridyl-N-based the electrochemical detection of small organic antigens in the health care area.

Inactivation of Castor Bean Allergen CB-1A by Heating and Chemical Treatment

  • Kim, Byong-Ki
    • Food Science and Biotechnology
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    • 제15권3호
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    • pp.441-446
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    • 2006
  • The biological effects of heating and chemical treatment on castor meal were investigated in order to develop a procedure to inactivate its antigenic activity in a way that is suitable for industrial applications. A 1% solution of purified castor bean allergen (CB-1A) was heat-treated with or without exposure to NaOH and NaOCI (250 ppm each). CB-1A exhibited extreme stability when heat-treated alone. In the presence of NaOH and NaOCl, CB-1A showed a drastic decrease in antigenic activity as the temperature surpassed the critical level of $70^{\circ}C$. The gradual disappearance of disc gel electrophoresis bands presumably responsible for the allergenicity of CB-1A, along with the significant losses of the amino acids phenylalanine, methionine, arginine, histidine, and cysteine correlated with the loss of CB-1A activity. CB-1A showed a single symmetrical band in SDS acrylamide gel electrophoresis with an estimated molecular weight of 6,000 daltons. The chemical and heat treatments reduced the disulfide bond content of CB-1A by 9.1% with a coincident increase in sulfhydryl bonds.

Surface Plasmon Resonance Immunosensor for Detection of Legionella pneumophila

  • Oh, Byung-Keun;Lee, Woochang;Bae, Young-Min;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.112-116
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    • 2003
  • An immunosensor based on surface plasmon resonance (SPR) onto a protein G layer by Self-assembly technique was developed for detection of Legionella pneumophila. The protein G layer by self-assembly technique was fabricated on a gold (Au) surface by adsorbing the 11-mercaptoundecanoic acid (MUA) and an activation process for the chemical binding of the free amino (-NH$_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of the protein G layer by self-assembly technique on the Au Substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The Surface topographies of the fabricated thin films on an Au substrate were also analyzed by using an atomic force microscope (AFM). Consequently, an immunosensor for the detection of L. pneumophila using SPR was developed with a detection limit of up to 10$^2$CFU per mL.

Detection of Escherichia coli O157:H7 Using Immunosensor Based on Surface Plasmon Resonance

  • Oh, Byung-Keun;Kim, Young-Kee;Bae, Young-Min;Lee, Won-Hong;Choi, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • 제12권5호
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    • pp.780-786
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    • 2002
  • An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

Yeast cell surface display of cellobiohydrolase I

  • Lee, Sun-Kyoung;Suh, Chang-Woo;Hwang, Sun-Duk;Kang, Whan-Koo;Lee, Eun-Kyu
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.468-472
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    • 2003
  • Recently, genetic engineering techniques have been used to display various heterologous peptides and proteins (enzyme, antibody, antigen, receptor and fluorescence protein, etc.) on the yeast cell surface. Living cells displaying various enzymes on their surface could be used repeatedly as 'whole cell biocatalysts' like immobilized enzymes. We constructed a yeast based whole cell biocatalyst displaying T. reesei cellobiohydrolase I (CBH I ) on the cell surface and endowed the yeast-cells with the ability to degrade cellulose. By using a cell surface engineering system based on ${\alpha}-agglutinin,$ CBH I was displayed on the cell surface as a fusion protein containing the N-terminal leader peptide encoding a Gly-Ser linker and the $Xpress^{TM}$ epitope. Localization of the fusion protein on the cell surface was confirmed by confocal microscopy. In this study, we report on the genetic immobilization of T. reesei CBH I on the S. cerevisiae and hydrolytic activity of cell surface displayed CBH I.

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