• Title/Summary/Keyword: Immunoassays

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Simultaneous Dual-Enzyme Immunoassays in a Solid Phase

  • 백세환;박순재
    • Bulletin of the Korean Chemical Society
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    • v.18 no.1
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    • pp.44-50
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    • 1997
  • A method of dual-signal generation from two different enzymes was developed and utilized to simultaneously perform dual immunoassays in a single microwell. Two enzymes selected as tracers were horseradish peroxidase (HRP) and β-galactosidase (GAL). 3, 3', 5, 5'-Tetramethylbenzidine (TMB) and chlorophenolred-β-galactopyranoside (CPRG) as chromogenic substrates for the respective enzyme were used. Although the two enzymes showed their maximum activities at distinct pH conditions (pH 5.1 for HRP and 7.5 for GAL), the enzyme reactions were able to be concurrently carried out at pH 5.75 in a dual-substrate solution without signal loss. This performance was achieved by increasing TMB concentration two-fold, introducing potassium salt as activator of GAL reaction, and extending total reaction time 50%. The signal generation method was then used for dual-enzyme immunoassays to detect antibodies with co-immobilized Hepatitis C virus antigens (core and NS5) and a Hepatitis B virus antigen (PreS(2)) in a microwell. Dose-response curves of the assays revealed cooperativity between different antigen-antibody complex formation, which suggested that dual immunoassays can only be used for qualitative screening tests unless the antigens immobilized were spatially separated.

Cross-reactivity of Amphetamine Analogues in Various Immunoassays (면역분석법에서 암페타민류의 교차 반응성)

  • 박미정;최화경;최상길;손행자;임미애;정희선
    • YAKHAK HOEJI
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    • v.47 no.5
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    • pp.266-270
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    • 2003
  • We evaluated four commercially available methamphetamine immunoassays for their relative cross-reactivities of amphetamine analogues in human urine: Abbott TDx, Vitalab Selectra and on-site test kits (Accusign MET, SD bioline MET). High cross-reactivities were shown at designer's drugs such as methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA) and methylenedioxyethylamphetamine (MDEA) in all of the tested immunoassays. Methoxyphenamine, fenfluramine and phentermine were positive in TDx and Selectra, but were not positive in on-site test kits. Pseudoephedrine, norpseudoephedrine, ephedrine, norephedrine, MDMA, MDA, fenfluramine and phentermine were detected by gas chromatography/mass spectrometry(GC/MS) in false positive urines. Since the overall specificity of any of the devices was not 100%, we found it is important to confirm any positive screening test result, so we developed simultaneous determination of amphetamine analogues in urines. After alkalinization of the urine samples with 6-N NaOH, the analytes were extracted using ethyl acetate, derivatized with pentafluoropropyl anhydride (PFPA) prior at GC/MS analysis.

Screening and Confirmation of Designer Drugs and Anorectics in Urines using Immunoassay and GC/MS

  • Choi, Hwa-Kyung;Park, Mee-Jung;Choi, Sang-Gil;Son, Haeng-Ja;Chung, Hee-Sun
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.297.1-297.1
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    • 2002
  • Immunoassays are frequently used for an screening method to detect the presence of drugs in Urine. The main advantages of the method are well known -- simplicity of handling samples. rapidity. sensitivity. and specificity of analysis. However. it is also known that immunoassays exhibit cross-reactivity to related drugs and there are only limited specific immunoassays on the market. This study reports on the ability of TDx to detect urine samples obtained from suspects of taking over-the-counter medications and illegal drugs containing ATS. designer drugs. (omitted)

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Immunoassay for Monitoring Pesticide Contamination in Agricultural Products

  • Park, Eun-Kee;Lee, Hu-Jang
    • Journal of Environmental Health Sciences
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    • v.34 no.6
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    • pp.433-438
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    • 2008
  • Much of the increase in agricultural productivity over the past half century has been due to the control of the pests with synthetic pesticides. The use of these pesticides has caused environmental problems and public health concern. The guidelines of maximum residue levels of pesticides in agricultural products has been well documented but more careful monitoring of their residues is required. Pyrethorid class pesticides are dominant in modern agricultural industry but public health concerns have been recently considered. The major route of pesticide exposure is the diet and with improved surveillance of pyrethorid residues in agricultural products their exposure should be controlled and minimized. In suitable products with reduced matrix effects such as agricultural products, aqueous samples, fruits and vegetables the use of immunoassays for pyrethorid residue monitoring could satisfy this requirement. Immunoassays have several advantages, namely they are highly sensitive, selective and cost-effective and enable large-scale sample handling and analysis in the laboratory.

Simultaneous Quantitative Determination of Multiple Analytes with Fluorescence- Tagged Probes by Immunochromatogratphy

  • Jeong, Dong-Seok;Choi, Eui-Yul
    • Animal cells and systems
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    • v.7 no.1
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    • pp.89-92
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    • 2003
  • Immunoassays have become indispensable tools and achieved great importance in scientific and medical research. However, typical immunoassays are time-consuming and use complex, multi-step procedures. In this study, we introduce a new immunoassay system for the quantification of several analytes at a time without any washing steps. It is comprised of a detector solution with fluorescence-labeled antibodies and a test strip with immobilized capture antibodies. Using a micro-array scanner, the antigen-antibody complex was quantitatively determined by measuring the intensities of fluorescence on the capture lines or dots of nitrocellulose membrane. This method demonstrated its rapid quantitative determination of analytes without many processing steps as well as specific identification of multiple analytes in biological specimens.

Developments of Hormone Assays, Second Generation: Non-Isotopic Immunoassays (호르몬 측정법의 발달 제 2세대: 비방사면역측정법)

  • Lee, Chang-Joo;Kim, Sang Soo;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.9 no.2
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    • pp.65-83
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    • 2005
  • The three important phases in the development of ligand immunoassays are identified and summarized. The competitive radiolabelled hormone measurement had been developed in the first and early in the second generations(1950s to 1960s), such as radioimmunoassays(RIA) or immunoradiometric(saturation) assays(IRMA), and used in all most of the hormone and also analyte in biological samples. In the second generation, ultrasensitive non-isotopic immunoassays(NIA) were developed using monoclonal antibodies(McAb), labelling the McAb and high specific activity non-isotopic labels. After their usefulness, advantages and disadvantages has been evaluated and non-competitive methods are discussed. The chip/microarray based multianalyte ligand assays(microspot or genechip methods) are developed and known as alternative ones in the third generation. We summarize the developments of NIAs and its usefulness, and then introduce briefly the new ligand assays.

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