Novel biotinylated acridinium derivatives: new reagents for fluorescence immunoassays and proteomics. 2005

Andreas Scorilas, and Kalomoira Agiamarnioti, and Kyriakos Papadopoulos
Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, 15701 Athens, Greece. ascorilas@biol.uoa.gr

BACKGROUND The favorable properties of acridinium derivatives over conventional fluorescent probes have attracted considerable interest. These conjugates have found wide utility in tests developed for medical diagnosis providing at least comparable sensitivity to radioimmunoassay (RIA) and enzyme immunoassay (EIA) procedures. METHODS In this paper we describe the synthesis, luminescent properties and immunoassay applications of two novel biotinylated acridinium derivatives, 9-(2-biotinyl-oxyethyl)-carboxylate-10-methyl-acridinium triflate, BOCMAT and 9-(2-biotinyl-amidoethyl)-carboxylate-10-methyl-acridinium triflate, BACMAT. RESULTS The chemiluminescence efficiency of the novel reagents is high in polar aprotic solvents and attains detection limits down to 7.28 x 10(-8) M, while the fluorescence efficiency has its maximum in aqueous solutions with detection limits down to 1.94 x 10(-10) M. The above novel compounds were applied in the fluorimetric determination of solid phase-immobilized biotinylated mouse IgG and the detection limit was found to be approximately 1 microg/assay ( approximately 7 pmol/assay). CONCLUSIONS The new streptavidin-based detection reagents were synthesized can be used for the development of highly sensitive solid-phase fluorescence-based immunoassays and may find important applications in proteomics.

UI MeSH Term Description Entries
D007074 Immunoglobulin G The major immunoglobulin isotype class in normal human serum. There are several isotype subclasses of IgG, for example, IgG1, IgG2A, and IgG2B. Gamma Globulin, 7S,IgG,IgG Antibody,Allerglobuline,IgG(T),IgG1,IgG2,IgG2A,IgG2B,IgG3,IgG4,Immunoglobulin GT,Polyglobin,7S Gamma Globulin,Antibody, IgG,GT, Immunoglobulin
D002138 Calibration Determination, by measurement or comparison with a standard, of the correct value of each scale reading on a meter or other measuring instrument; or determination of the settings of a control device that correspond to particular values of voltage, current, frequency or other output. Calibrations
D005453 Fluorescence The property of emitting radiation while being irradiated. The radiation emitted is usually of longer wavelength than that incident or absorbed, e.g., a substance can be irradiated with invisible radiation and emit visible light. X-ray fluorescence is used in diagnosis.
D000166 Acridines Compounds that include the structure of acridine. Acridine
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001710 Biotin A water-soluble, enzyme co-factor present in minute amounts in every living cell. It occurs mainly bound to proteins or polypeptides and is abundant in liver, kidney, pancreas, yeast, and milk. Vitamin H,Biodermatin,Biokur,Biotin Gelfert,Biotin Hermes,Biotin-Ratiopharm,Biotine Roche,Deacura,Gabunat,Medebiotin,Medobiotin,Rombellin,Biotin Ratiopharm,Gelfert, Biotin,Hermes, Biotin,Roche, Biotine
D015200 Fluoroimmunoassay The use of fluorescence spectrometry to obtain quantitative results for the FLUORESCENT ANTIBODY TECHNIQUE. One advantage over the other methods (e.g., radioimmunoassay) is its extreme sensitivity, with a detection limit on the order of tenths of microgram/liter. Immunofluorometric Assay,Time-Resolved Immunofluorometric Assay,Assay, Immunofluorometric,Assay, Time-Resolved Immunofluorometric,Assays, Immunofluorometric,Assays, Time-Resolved Immunofluorometric,Fluoroimmunoassays,Immunofluorometric Assay, Time-Resolved,Immunofluorometric Assays,Immunofluorometric Assays, Time-Resolved,Time Resolved Immunofluorometric Assay,Time-Resolved Immunofluorometric Assays
D015394 Molecular Structure The location of the atoms, groups or ions relative to one another in a molecule, as well as the number, type and location of covalent bonds. Structure, Molecular,Molecular Structures,Structures, Molecular
D049449 Luminescence Emission of LIGHT when ELECTRONS return to the electronic ground state from an excited state and lose the energy as PHOTONS. It is sometimes called cool light in contrast to INCANDESCENCE. LUMINESCENT MEASUREMENTS take advantage of this type of light emitted from LUMINESCENT AGENTS. Luminescence, Physical,Chemiluminescence,Chemiluminescence, Physical,Physical Chemiluminescence,Physical Luminescence
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

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