• 제목/요약/키워드: sandwich-type immunoassay

검색결과 8건 처리시간 0.02초

Bioanalytical Application of SERS Immunoassay for Detection of Prostate-Specific Antigen

  • Yoon, Kyung-Jin;Seo, Hyeong-Kuyn;Hwang, Hoon;Pyo, Dong-Jin;Eom, In-Yong;Hahn, Jong-Hoon;Jung, Young-Mee
    • Bulletin of the Korean Chemical Society
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    • 제31권5호
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    • pp.1215-1218
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    • 2010
  • We demonstrate the possible application of the sandwich type surface-enhanced Raman scattering (SERS) immunoassay using antigen-antibody binding for detection of prostate-specific antigen (PSA) in cancer cells. In this sandwich type of SERS immunoassay, to capture antigens onto the immobilized layer of antibodies on the gold substrate we prepared the monolayer of gold nanoparticles on the APTMS-derivatized surface of a glass slide by using the SAM technique. This sandwich type of SERS immunoassay in which antigens on the substrate specifically capture antibodies on a Raman reporter (DSNB coated gold nanoparticles with R6G) could successfully detect PSA at low levels. A strong SERS spectrum of Raman reporter was observed only with a substrate in which PSA is present.

Evaluation of Two Types of Biosensors for Immunoassay of Botulinum Toxin

  • Choi, Ki-Bong;Seo, Won-Jun;Cha, Seung-Hee;Choi, Jung-Do
    • BMB Reports
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    • 제31권1호
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    • pp.101-105
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    • 1998
  • Immunoassay of botulinum toxin (BTX) B type was investigated using two typed of biosensors: light addressable potentiometric sensor (LAPS) and surface plasmon resonance (SPR) sensor. Urease-tagged and immuno-filtration capture method have been used for LAPS. Tag-free and direct binding real-time detection method have been used for SPR sensor. The detection limit of sandwich assay format with LAPS was 10 ng/ml, which was the lowest among methods tested. SPR has the advantage of being more convenient because tag-free direct binding assay can be used and reaction time was reduced, regardless of low sensitivity. This result shows that sandwich assay format with LAPS can be used as an alternative method of BTX mouse bioassay which is known as the most sensitive method for the detection of BTX.

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Protein Array Fabricated by Microcontact Printing for Miniaturized Immunoassay

  • Lee Woo-Chang;Lim Sang-Soo;Choi Bum-Kyoo;Choi Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • 제16권8호
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    • pp.1216-1221
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    • 2006
  • A protein array was fabricated for a miniaturized immunoassay using microcontact printing ($\mu$CP). A polydimethylsiloxane (PDMS) stamp with a 5 $\mu$m$\times$5 /$\mu$m dimension was molded from a silicon master developed by photolithography. Under optimal fabrication conditions, including the baking, incubation, and exposure time, a silicon master was successfully fabricated with a definite aspect ratio. An antibody fragment was utilized as the ink for the $\mu$CP, and transferred to an Au substrate because of the Au-thiol (-SH) interaction. The immobilization and antibody-antigen interaction were investigated with fluorescence microscopy. When human serum albumin (HSA) was applied to the protein array fabricated with an antibody against HSA, the detection limit was 100 pg/ml of HSA when using a secondary antibody labeled with a fluorescence tag. The fabricated protein array maintained its activity for 14 days.

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

  • 차상훈;김희주;김원배;양중익
    • 약학회지
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    • 제31권2호
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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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Human Cardiac Troponin I 면역분석을 위한 단일클론 항체의 특성화

  • 오흥일;양진아;백의환;백세환
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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    • pp.713-714
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    • 2000
  • Six monoclonal antibodies to human cardiac troponin I (hcTnI) were produced to eventually develop an immunosensor for acute myocardial infarction (AMI). For the characterization of these antibodies, a set of 11 different peptides covering selected ranges of the complete amino acid sequence of hcTnI was prepared and used for epitope mapping. Such analysis allowed to select an appropriate pair of antibodies that can form a sandwich type of immune complexes and was consequently used for an immunoassay.

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식육중 Escherichia coli O157 검출을 위한 enzyme immunoassay 기법 개발 (Development of an enzyme immunoassay for detection of Escherichia coli O157 in meat)

  • 정병열;정석찬;조동희;김종염;박용호;신쌍재;김성국;김봉환
    • 대한수의학회지
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    • 제38권4호
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    • pp.745-750
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    • 1998
  • A sensitive and rapid enzyme immunoassay(EIA) to detect Escherichia coli O157 in ground beef was developed by using a sandwich type assay with polyclonal antibodies to E coli O157. E coli O157 in ground beef could be detected within 15hr, including incubation for 12hr in enrichment broth and 3hr in immunoassay. The EIA could detect $1.3{\times}10^5$ cells of E coli O157/g of ground beef without enrichment. The lowest limit of detection was 0.23 E coli O157 per g of meat after enrichment. Confirmation was required in the positive specimens in the EIA by culture method even though the negative specimens were not. These results suggested that the immunoassay could be a very efficient method for the screening E coli O157 in meat.

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A New Concept for Efficient Sensitivity Amplification of a QCM Based Immunosensor for TNF-α by Using Modified Magnetic Particles under Applied Magnetic Field

  • Bahk, Yeon-Kyoung;Kim, Hyung-Hoon;Park, Deog-Su;Chang, Seung-Cheol;Go, Jeung-Sang
    • Bulletin of the Korean Chemical Society
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    • 제32권12호
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    • pp.4215-4220
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    • 2011
  • This study introduces a new concept for a simple, efficient and cheap sensitivity amplification of a Quartz Crystal Microbalance (QCM) based immunosensor system for the detection of tumor necrosis factor-alpha (TNF-${\alpha}$, TNF) by using an in-built magnetic system. The frequency shift due to the applied magnetic field was successfully observed on magnetic particles labeled detection antibodies, anti-human TNF-${\alpha}$, which were bound to the immunologically captured TNF-${\alpha}$ on the gold coated quartz crystals. In the present system, the magnitude of frequency shift depends on both the strength of magnetic field and the amount of target antigen applied. Significant signal amplification was observed when the additional built-in residual stress generated by the modified magnetic particles under the magnetic field applied. Used in conjunction with a sandwich type non-competitive immunoassay format, the lower detection limit was calculated to be 25 $ngmL^{-1}$ and showed good linearity up to TNF-${\alpha}$ concentrations as high as 2.0 ${\mu}gmL^{-1}$. The sensitivity, most importantly, was improved up to 4.3 times compared with the same QCM system which was used only an antigen-antibody binding without additional magnetic amplification.

Development of Disposable Immunosensors for Rapid Determination of Sildenafil and Vardenafil in Functional Foods

  • Vijayaraj, Kathiresan;Lee, Jun Hyuck;Kim, Hyung Sik;Chang, Seung-Cheol
    • 한국식품위생안전성학회지
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    • 제32권2호
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    • pp.83-88
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    • 2017
  • 본 연구의 목적은 screen printed 탄소계 전극에 기반하여 기능성 식품중에 포함된 소량의 Sildenafil과 Vardenafil(SDF 및 VDF)을 쉽게 검출하기 위한 전류계 면역센서를 개발하고자 하였다. 본 연구에서 개발한 면역센서는 horseradish peroxidase로 labeling 시킨 후 비경쟁적 샌드위치 ELISA 측정법에 의한 원리를 이용하였다. 개발된 센서는 그래핀 산화물 및 키토산(ErGO-CS) 복합체를 단순한 전기화학적 증착에 의해 screen printed 탄소계 전극을 이용하는 원리이며, 감지된 화학물질을 평가하고 센서특성을 최적화하기 위해 전기 화학적 임피던스분광법, 순환 전압 전류법 등을 포함한 일련의 전기화학적 실험방법에 기초하여 완성되었다. 본 연구에서 개발된 센서는 100 pg/mL ~ 300 ng/mL 농도의 SDF 및 VDF에 대하여 직선상의 농도-의존적인 반응을 나타내었다. 또한 최저 검출 한계는 55 pg/mL이었으며, 센서의 민감도는 $1.02{\mu}Ang/mL/cm^2$로 산출되었다. 센서의 성능은 7.1%의 상대 표준편차로 매우 우수한 재현성을 나타내었다. 본 연구에서 개발된 센서의 전략적 가치는 향후 건강기능식품중에 함유된 SDF 및 VDF과 같은 의약품의 미량을 현장에서 쉽게 분석 할 수 있어서 비용-효과적 측면에서 그 가치가 우수하다는 것을 제시한다.