Novel biotinylated acridinium derivatives: new reagents for fluorescence immunoassays and proteomics.
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.
Scorilas
Andreas
A
Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, 15701 Athens, Greece. ascorilas@biol.uoa.gr
Agiamarnioti
Kalomoira
K
Papadopoulos
Kyriakos
K
D000166:Acridines / Q000138:chemical synthesis* / Q000737:chemistry; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D019921:Biotinylation; D002138:Calibration; D005453:Fluorescence; D015200:Fluoroimmunoassay / Q000295:instrumentation* / Q000379:methods*; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D049449:Luminescence; D051379:Mice; D015394:Molecular Structure; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D019809:Streptavidin / Q000737:chemistry
C504967:9-(2-biotinylamidoethyl)carboxylate-10-methylacridinium; C504966:9-(2-biotinyloxyethyl)carboxylate-10-methylacridinium; D000166:Acridines; D007074:Immunoglobulin G; D001710:Biotin; D019809:Streptavidin