Fluorescence immunoassay enzymatic

T. K. Christopoulos and E. P. Diamandis, Enzymatically amplified time-resolved fluorescence immunoassay with terbium chelates, Anal. Chem. 64, 342-346 (1992). 

The hydrolysis of a nonfluorescent enzyme substrate to a fluorescent product is widely utilized to measure the activity of a large number of enzymes. Binding of enzyme substrates by antibodies often protects the enzymatically labile bond from hydrolysis. By the combination of these two formats, the substrate-labeled fluorescent immunoassay (SLFIA) was developed. ... 

Figure 6.3. Substrate-labeled fluorescent immunoassay for theophylline. (A) Effect of theophylline rabbit antiserum (O) and normal rabbit serum (A) on the enzymatic hydrolysis of 8-[3-(7-/l-galactosylcoumarin-3-carboxyamido)propyl]theophylline. (B) Effect of various concentrations of theophylline on the fluorescence resulting from enzymatic hydrolysis of 8-[3-(7-/)-galactosyl-coumarin-3-carboxyamido)propyI]theophylline. (Reprinted from Ref. 3, with permission from the American Association for Clinical Chemistry.)...

Figure 6.4. Schematic representation of a fluorescent immunoassay for theophylline utilizing enzymatic hydrolysis of an intramolecularly quenched theophylline conjugate of flavin adenine dinucleotide. (Reprinted from Ref. 5, with permission from Academic Press.)...

T. A. Kelly and G. D. Christian, Homogeneous Enzymatic Fluorescence Immunoassay of Serum IgG by Continuous Flow Injection Analysis. Ta-lanta, 29 (1982) 1109. 

Selected entries from Methods in Enzymology [vol, page(s)] Design, 178, 551 immunoassay, 178, 542 production, 178, 531 purification, 178, 543 substrates and enzymatic assay, 178, 544 derivatization with spectroscopic probe, 178, 567 ester cleavage assays, 178, 565 fluorescence quenching binding assay, 178,... 

Sensitivity of immunoassays is largely conditioned by markers applied for conjugation with the antibody. Traditional immunodetection methods ELISA with radioactive markers (radioimmunoassay—RIA), enzymatic markers (enzyme immunoassays—El A), or fluorescent markers (fluoroenzyme immunoassays FEIA) are currently the most widely used techniques in laboratory analysis of allergens as well as in clinical studies for determination of general and specific IgE and other subclasses of immunoglobulines, e.g., IgG4 in the Immuno-CAP system (Samson, 2001 Duran-Tauleria et al., 2004 Lidholm et al., 2006). 

The ELISA method (in its various modifications), employing radionuclide markers (radioimmunoassay RIA), enzymatic markers (enzyme immunoassay EIA) (Dill et al., 2006), or fluorescent dyes (fluorophores or fluorochromes) (Hildebrand et al., 2005) (FEIA—fluoroenzyme immunoassay, Immuno-CAP-system) or nanoparticles (Dequaire et al., 2000) are commonly used as materials for protein labeling in an immunoassay. 

Enzymatic Hydrolysis of Antigen-Fluorescent Dye Conjugate Immunoassay... 

Tsuji A (1981) Enzymatic assay by chemiluminescence and fluorescence reactions. In Ishikawa E, Kawai T, and Miyai K (eds.) Enzyme Immunoassay, pp. 41-53. Tokyo Igaku-Shoin. 

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