20 research outputs found
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A dissociative fluorescence enhancement technique for one-step time-resolved immunoassays
The limitation of current dissociative fluorescence enhancement techniques is that the lanthanide chelate structures used as molecular probes are not stable enough in one-step assays with high concentrations of complexones or metal ions in the reaction mixture since these substances interfere with lanthanide chelate conjugated to the detector molecule. Lanthanide chelates of diethylenetriaminepentaacetic acid (DTPA) are extremely stable, and we used EuDTPA derivatives conjugated to antibodies as tracers in one-step immunoassays containing high concentrations of complexones or metal ions. Enhancement solutions based on different ÎČ-diketones were developed and tested for their fluorescence-enhancing capability in immunoassays with EuDTPA-labelled antibodies. Characteristics tested were fluorescence intensity, analytical sensitivity, kinetics of complex formation and signal stability. Formation of fluorescent complexes is fast (5Â min) in the presented enhancement solution with EuDTPA probes withstanding strong complexones (ethylenediaminetetra acetate (EDTA) up to 100Â mM) or metal ions (up to 200Â ÎŒM) in the reaction mixture, the signal is intensive, stable for 4Â h and the analytical sensitivity with Eu is 40Â fmol/L, Tb 130Â fmol/L, Sm 2.1Â pmol/L and Dy 8.5Â pmol/L. With the improved fluorescence enhancement technique, EDTA and citrate plasma samples as well as samples containing relatively high concentrations of metal ions can be analysed using a one-step immunoassay format also at elevated temperatures. It facilitates four-plexing, is based on one chelate structure for detector molecule labelling and is suitable for immunoassays due to the wide dynamic range and the analytical sensitivity
Time-Resolved FRET -Based Approach for Antibody Detection - A New Serodiagnostic Concept
Peer reviewe
Homogeneous Assay Based on Anti-Stokes' Shift Time-Resolved Fluorescence Resonance Energy-Transfer Measurement
Influence of Coupling Method on the Luminescence Properties, Coupling Efficiency, and Binding Affinity of Antibodies Labeled with Europium(III) Chelates
Fluorescence Quenching-Based Assays for Hydrolyzing Enzymes. Application of Time-Resolved Fluorometry in Assays for Caspase, Helicase, and Phosphatase
A New Simple Cell-Based Homogeneous Time-Resolved Fluorescence QRET Technique for Receptor-Ligand Interaction Screening
Time-resolved lanthanide luminescence for lab-on-a-chip detection of biomarkers on cancerous tissues
PDMS-based microfluidic devices combined with lanthanide-based immunocomplexes have been successfully tested for the multiplex detection of biomarkers on cancerous tissues, revealing an enhanced sensitivity compared to classical organic dyes