• Title/Summary/Keyword: Electrochemical immunoassay

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An Electrochemical Enzyme Immunochip Based on Capacitance Measurement for the Detection of IgG

  • Yi, Seung-Jae;Choi, Ji-Hye;Kim, Hwa-Jung;Chang, Seung-Cheol;Park, Deog-Su;Kim, Kyung-Chun;Chang, Chulhun L.
    • Bulletin of the Korean Chemical Society
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    • v.32 no.4
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    • pp.1298-1302
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    • 2011
  • This study describes the development of an electrochemical array immunochip for the detection of IgG. Interdigitated immunochip platforms were fabricated by sputtering gold on a glass wafer by using MEMS process and then were coated with Eudragit S100, an enteric polymer, forming an insulating layer over the working area of immunochips. The breakdown of the polymer layer was exemplified by the catalytic action of urease which, in the presence of urea, caused an alkaline pH change. This subsequently caused an increase of the double layer capacitance of the underlying electrode. Used in conjunction with a competitive immunoassay format, this allowed the ratio of initial to final electrode capacitance to be directly linked with the concentration of analyte, i.e. IgG. Responses to IgG could be detected at IgG concentration as low as $250\;ngmL^{-1}$ and showed good linearity up to IgG concentration as high as $20\;{\mu}gmL^{-1}$.

Electrochemical Immunosensing of GOx-labeled CRP Antigen on Capture Antibody Monolayer Immobilized on Calixcrown-5 SAMs

  • Jung, Hye-Sook;Song, Kum-Soo;Kim, Tai-Sun
    • Bulletin of the Korean Chemical Society
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    • v.28 no.10
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    • pp.1792-1796
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    • 2007
  • Insulating effects on Au electrode according to the thickness and density of coated materials are well-known. To do electrochemical immunoassay reproducibly the glod electrode would be coated with self-assembled monolayers and antobodies. To get reproducibility, the antobody monolayer should be packed at highest density so that the amount of immobilized antibody at defined area should be the same. The calix[4]crown-5 SAMs could provide the basis for the antibodies to be immobilized reproducibly and at highest density. But the insulating effect would be highest too. We proved that the compactly packed protein monolayers on SAMs inhibited the electron transfer by block the free shuttling of redox molecules. The inhibition was minimized by inserting redox molecules in between the proteins during immobilization process. In this paper, we demonstrated that the calix[4]crown-5 SAMs would provide the protein monolayers with highest density and new method to minimize the insulating effect by inserted redox molecules in between the compactly packed protein monolayers.

One-Touch Type Immunosenging Lab-on-a-chip for Portable Point-of-care System (휴대용 POC 시스템을 위한 원터치형 면역 센싱 랩온어칩)

  • Park, Sin-Wook;Kang, Tae-Ho;Lee, Jun-Hwang;Yoon, Hyun-C.;Yang, Sang-Sik
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.56 no.8
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    • pp.1424-1429
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    • 2007
  • This paper presents a simple and reliable one-touch type multi-immunosensing lab-on-a-chip (LOC) detecting antibodies as multi-disease markers using electrochemical method suitable for a portable point-of-care system (POCS). The multi-stacked LOC consists of a PDMS space layer for liquids loading, a PDMS valve layer with 50 im in height for the membrane, a PDMS channel layer for the fluid paths, and a glass layer for multi electrodes. For the disposable immunoassay which needs sequential flow control of sample and buffer liquids according to the designed strategies, reliable and easy-controlled on-chip operation mechanisms without any electric power are necessary. The driving forces of sequential liquids transfer are the capillary attraction force and the pneumatic pressure generated by air bladder push. These passive fluid transport mechanisms are suitable for single-use LOC module. Prior to the application of detection of the antibody as a disease marker, the model experiments were performed with anti-DNP antibody and anti-biotin antibody as target analytes. The flow test results demonstrate that we can control the fluid flow easily by using the capillary stop valve and the PDMS check valves. By the model tests, we confirmed that the proposed LOC is easily applicable to the bioanalytic immunosensors using bioelectrocatalysis.

Mass Spectrometry-Based Analytical Methods of Amatoxins in Biological Fluids to Monitor Amatoxin-Induced Mushroom Poisoning

  • Choi, Jin-Sung;Lee, Hye Suk
    • Mass Spectrometry Letters
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    • v.13 no.4
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    • pp.95-105
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    • 2022
  • Amatoxin-induced mushroom poisoning starts with nonspecific symptoms of toxicity but hepatic damage may follow, resulting in the rapid development of liver insufficiency and, ultimately, coma and death. Accurate detection of amatoxins, such as α-, β-, and γ-amanitin, within the first few hours after presentation is necessary to improve the therapeutic outcomes of patients. Therefore, analytical methods for the identification and quantification of α-, β-, and γ-amanitin in biological samples are necessary for clinical and forensic toxicology. This study presents a literature review of the analytical techniques available for amatoxin detection in biological matrices, and established an inventory of liquid chromatography (LC) techniques with mass spectrometry (MS), ultraviolet (UV) detection, and electrochemical detection (ECD). LC-MS methods using quadrupole tandem mass spectrometry, time-of-flight mass spectrometry, and orbitrap MS are powerful analytical techniques for the identification and determination of amatoxins in plasma, urine, serum, and tissue samples, with high sensitivity, specificity, and reproducibility compared to LC with UV and ECD, enzyme-linked immunoassay, and capillary electrophoresis methods.

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
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    • v.32 no.2
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    • pp.83-88
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    • 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.

ANALYSIS OF FLUIDIC BEAD CUBE EMBEDDED PORTABLE CMOS SENSING SYSTEM FOR IMMUNO REACTION MONITORING (유체소자가 집적화된 면역검사용 휴대용 CMOS 바이오칩의 분석)

  • Jeong, Yong-Won;Park, Se-Wan;Kim, Jin-Seok;Kim, Hyeon-Cheol;Chun, Kuk-Jin
    • Proceedings of the IEEK Conference
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    • 2005.11a
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    • pp.755-758
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    • 2005
  • This paper describes the novel immunoassay sensing system for a portable clinical diagnosis system. It consists of a bead cage reactor and a CMOS integrated biosensor. It showed the simple and easy antibody coating method on beads by flow-through avidin biotin complex technology in a microfluidic device. It showed just 90 nL sample consumption and good result for the application of alpha feto protein. The bead cage reactor has the role of the antibody coating, antigen binding and enzyme linking for the electrochemical sensing method. The CMOS biosensor consists of ISFET (ion selective field effect transistor) biosensor and temperature sensor for detecting pH that is the byproduct of enzyme reaction. The sensitivity is 8 $kHz/^{\circ}C$ in a temperature sensor and 33 mV/pH in a pH sensor. After filling the 15 um polystyrene beads in bead cage, antibody flowed and reacted to beads. Subsequently, the biotinylated antigen flowed and bound to the antibody and GOD (glucose oxidase)-avidin conjugate flowed and reacted to the biotin of the biotinylated antigen. After this reaction process, glucose solution flowed and reacted to the GOD on beads. The hydrogen was generated by glucose-GOD reaction. And it was detected by the pH sensor.

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Research Trend of Biochip Sensors for Biomarkers Specific to Diagnostics of Lung Cancer Diseases (폐암 질환 진단에 활용 가능한 바이오마커 검출용 바이오칩 센서 연구 동향)

  • Lee, Sang Hyuk;Goh, Eunseo;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.29 no.6
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    • pp.645-651
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    • 2018
  • Lung cancer has the highest death rate of any cancer diseases in Koreans. However, patients often feel difficult to recognize their disease before facing the terminal diagnosis due to the absence of any significant symptoms. Furthermore, the clear detection of an early cancer stage is usually obscure with existing diagnostic methods. For this reason, extensive research efforts have been made on introducing a wide range of biochemical diagnostic tools for the molecular level analysis of biological fluids for lung cancer diagnoses. A chip-based biosensor, one type of the analytical devices, can be a great potential for the diagnosis, which can be used without any further expensive analytical equipments nor skilled analysts. In this mini review, we highlight recent research trends on searching biomarker candidates and bio-chip sensors for lung cancer diagnosis in addition to discussing their future aspects.