• Title/Summary/Keyword: sandwich-type immunoassay

Search Result 8, Processing Time 0.027 seconds

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
    • /
    • v.31 no.5
    • /
    • pp.1215-1218
    • /
    • 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
    • /
    • v.31 no.1
    • /
    • pp.101-105
    • /
    • 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.

  • PDF

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
    • /
    • v.16 no.8
    • /
    • pp.1216-1221
    • /
    • 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.

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

  • 차상훈;김희주;김원배;양중익
    • YAKHAK HOEJI
    • /
    • v.31 no.2
    • /
    • pp.64-69
    • /
    • 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.

  • PDF

Human Cardiac Troponin I 면역분석을 위한 단일클론 항체의 특성화

  • O, Heung-Il;Yang, Jin-A;Baek, Ui-Hwan;Baek, Se-Hwan
    • 한국생물공학회:학술대회논문집
    • /
    • 2000.11a
    • /
    • pp.713-714
    • /
    • 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.

  • PDF

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

  • Jung, Byeong-yeal;Jung, Suk-chan;Cho, Dong-hee;Kim, Jong-yeom;Park, Yong-ho;Shin, Sang-jae;Kim, Sung-guk;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
    • /
    • v.38 no.4
    • /
    • pp.745-750
    • /
    • 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.

  • PDF

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
    • /
    • v.32 no.12
    • /
    • pp.4215-4220
    • /
    • 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
    • Journal of Food Hygiene and Safety
    • /
    • v.32 no.2
    • /
    • pp.83-88
    • /
    • 2017
  • We introduced disposable amperometric immunosensors for the detection of Sildenafil and Vardenafil (SDF/VDF) based on screen printed carbon electrodes. The developed immunosensors were used as a non-competitive sandwich-type enzyme immunoassay with a horseradish peroxidase label. The sensors were constructed on screen printed carbon electrodes by the simple electrochemical deposition of a reduced graphene oxide and chitosan (ErGO-CS) composite. To evaluate the sensing chemistry and optimize the sensor characteristics, a series of electrochemical experiments were carried out including electrochemical impedance spectroscopy, cyclic voltammetry and amperometry. The sensors showed a linear response to SDF/VDF concentrations in a range from 100 pg/mL to 300 ng/mL. The lower detection limit was calculated to be 55 pg/mL, the sensitivity was calculated to be $1.02{\mu}Ang/mL/cm^2$, and the sensor performance exhibited good reproducibility with a relative standard deviation (RSD) of 7.1%. The proposed sensing chemistry strategy and the sensor format can be used as a simple, cost-effective, and feasible method for the in-field analysis of SDF/VDF in functional or health supplement food samples.