Fluoroimmunoassays

The performance of microwave-assisted decomposition of most difficult samples of organic and inorganic natures in combination with the microwave-assisted solution preconcentration is illustrated by sample preparation of carbon-containing matrices followed by atomic spectroscopy determination of noble metals. Microwave-assisted extraction of most dangerous contaminants, in particular, pesticides and polycyclic aromatic hydrocarbons, from soils have been developed and successfully used in combination with polarization fluoroimmunoassay (FPIA) and fluorescence detection. 

Several other techniques for have evolved for biochemical assays. In chapter 2 of this book, Omann and Sklar report on a method of fluoroimmunoassay where the bound and unbound antigen are separated by the quenching of fluorescence that accompanies antibody binding. Then, in chapter 3, Holl and Webb show how they achieved a sensitive measurement of nucleic acids by the enhancement in fluorescence that accompanies the binding of fluorescent dyes to nucleic acids. Chandler et al, also used fluorescence enhancement to monitor calcium mobility in neutrophil cells. 

Fig.7. A demonstration of fluoroimmunoassay. Green circles (Tb-cored dendrimer complex) and red circles (conventional Eu complex) corresponds to different assays...

UEHARA M, ARAI Y, WATANABE s and ADLERCREUTZ H (2000) Comparison of plasma and urinary phytoestrogens in Japanese and Finnish women hy time-resolved fluoroimmunoassay. Biofactors. 12 (1-4) 217-25. 

STUMPF K, UEHARA M, NURMI T, ADLERCREUTZ H (2000) Changes in time-resolved fluoroimmunoassay of plasma enterolactone, Analytical Biochemistry, 284, 153-7. 

Enzyme-Linked Immunoadsorbent Assay (NRL) Multiplexed Sandwich Fluoroimmunoassay (LLNL) Capillary Electrophoresis (LLNL)... 

The determination of theophylline in plasma can also be accomplished by various immunoassay techniques.66-67 Theophylline was also determined by a polarization fluoroimmunoassays but found to have a caffeine interference.88. In a more research oriented application, the interaction of caffeine with L-tryptophan was studied using h NMR with the results indicating that caffeine interacted with tryptophan in a 1 1 molar ratio through parallel stacking.69... 

Nowadays, antibodies are utilized in numerous immunoanalytical methods. Those widely used in practice, such as radioimmunoassays, fluoroimmunoassays and enzyme-linked immunosorbent assays (ELISA), require labelled reagents. Millions of ELISA tests for diagnostics of various diseases are daily performed in clinical laboratories. The detection of analytes by two-antibody "sandwich" ELISA, is schematically outlined in Figure 3. 

Goldman, E.R. et al. (2002b) Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays. Anal. Chem. 74, 841-847. 

Goldman, E.R., Mattoussi, H., Anderson, G.P., Medintz, I.L., and Mauro, J.M. (2005) Fluoroimmunoassays using antibody-conjugated quantum dots. Meth. Mol. Biol. 303, 19-34. 

Hemmila, I. (1985) Fluoroimmunoassays and immunofluorometric assays [Review]. Clin. Chem. 31, 359-370. 

Stahlberg, T., Markela, E., Mikola, H., Mottram, P., and Hemmila, I. (1993) Europium and samarium in time-resolved fluoroimmunoassays. American Laboratory December, 15-20. 

BMP-IA bacterial magnetic particle-based immunoassay FIIA flow-injection immunoassay PFIA polarization fluoroimmunoassay. b BDD12AC benzyldimethyldodecylammonium chloride DDAC didecyldimethylammonium chloride. 

Cummins CM, Koivunen ME, Stephanian A, Gee SJ, Hammock BD, Kennedy IM (2006) Application of europium(III) chelate-dyed nanoparticle labels in a competitive atrazine fluoroimmunoassay on an ITO waveguide. Biosens Bioelectron 21 1077-1085... 

Scheme 10 Schematic illustration of the CCP-amplified NP-based fluoroimmunoassay...

Wang Y, Liu B (2009) Conjugated polymer as a signal amplifier for novel silica nanoparticle-based fluoroimmunoassay. Biosens Bioelectron 24 3293-3298... 

Immunoassays based on fluorescence polarization have become very popular because, in contrast to radioimmunoassays, they require no steps to separate free and bound tracer. In a fluoroimmunoassay, the fluorescently labeled antigen is... 

D. J. Bucher, A. Mikhail, S. Popple, P. Graves, G. Meiklejohn, D. S. Hodes, K. Johansson, andP. E. Halonen, Rapid detection of Type A Influenza viruses with monoclonal antibodies to the M protein (Ml) by enzyme-linked immunosorbent assay and time-resolved fluoroimmunoassay, J. Clin. Microbiol. 29, 2484-2488 (1991). 

B. J. Tromberg, M. J. Sepaniak, T. VoDinh, and G. D. Griffin, Fiber-optic chemical sensors for competitive binding fluoroimmunoassay, Anal. Chem. 59, 1226-1230(1987). 

Enzymes can be linked to immunoassay reagents to amplify detection by the use of fluorogenic substrates. Enzyme-linked fluoroimmunoassays (ELFIAs) are very similar to photometric EIAs in format and workflow. EIAs are widely used, and many commercial ELFIA assays and systems are available/15 The most commonly used enzymes in ELFIAs are horseradish peroxidase, alkaline phosphatase, and fi-D-... 

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