File:Homogeneous competitive assays (FPIA, EMIT, LOCI, KIMS and CEDIA).png
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DescriptionHomogeneous competitive assays (FPIA, EMIT, LOCI, KIMS and CEDIA).png |
English: Homogeneous competitive assays (FPIA, EMIT, LOCI, KIMS and CEDIA). Original article caption: In fluorescence polarization immunoassay (FPIA) after incubation, the fluorescence polarization signal is measured without separation of bound from free labels. Free labeled analyte analog molecules are added to the sample, and it has a different Brownian motion than when the label is bound to a large antibody (Ab). When the analyte is present, there is competition for the binding to the Ab. If the labeled analyte is bound to the Ab molecule then the signal is generated, while when the labeled antigen is free in solution no signal is produced. Therefore, signal intensity is inversely proportional to the analyte concentration. Enzyme multiplied immunoassay technique (EMIT). Free analyte analog molecules labeled with an enzyme, e.g., glucose-6-phosphate dehydrogenase enzyme, are added to the test solutions to compete to the analyte to be tested. The active enzyme reduces NAD (no signal) to NADH (absorbs at 340), so that absorbance is monitored at 340 nm. When labeled analyte binds to the Ab, the enzyme becomes inactive, and so the signal is generated by the free label, and signal intensity is directly proportional to the analyte concentration. Luminescent oxygen channeling immunoassay (LOCI). The reaction mixture is irradiated to generate singlet oxygen species in microbeads coupled to the analyte. When bound to the respective Ab molecule, also coupled to another kind of bead, the analyte reacts with singlet oxygen and chemiluminescence signals are generated proportionally to the concentration of the analyte–Ab complex. Kinetic interaction of microparticle in solution (KIMS) and the conceptually similar particle enhanced turbidimetric inhibition immunoassay (PETINIA). In the absence of the analyte, free antibodies bind to drug microparticles conjugates to form aggregates that absorb in the visible range. Absorbance (or turbidimetry) is monitored and in presence of the analyte the Ab binds to the free analyte preventing microparticle aggregation; a reduction in absorbance is observed (signal is inversely proportional to analyte concentration). Cloned enzyme donor immunoassay (CEDIA). An enzyme (like beta-galactosidase) is genetically engineered into two inactive fragments: a small one called enzyme donor (ED) conjugated with the drug analog, and a larger fragment enzyme acceptor (EA): when the two fragments associate, the full enzyme converts a substrate into a cleaved colored product. If drug analyte molecules are present, they will compete with the ED-labeled drug in solution for the limited Ab sites, so that free ED-labeled drug analog will bind to EA generating a colorimetric signal directly proportional to the amount of analyte. |
Date | |
Source |
Sanavio B, Krol S (2015). "On the Slow Diffusion of Point-of-Care Systems in Therapeutic Drug Monitoring.". Front Bioeng Biotechnol 3: 20. DOI:10.3389/fbioe.2015.00020. PMID 25767794. PMC: 4341557. - License: "CC BY 4.0" |
Author |
Sanavio B, Krol S. Minor corrections by Mikael Häggström |
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Date/Time | Thumbnail | Dimensions | User | Comment | |
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current | 07:23, 29 April 2024 | 3,369 × 1,673 (2.27 MB) | imagescommonswiki>Mikael Häggström | Juxtaposed similar asssays |
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