Simultaneous antigen detection using multiplex dyes

Galla, K.; Arden‐Jacob, Jutta; Deltau, Gerhard; Drexhage, K. H.; Martin, Michael; Sauer, Markus; Wolfrum, J.; Seeger, Stefan

doi:10.1007/bf01876665

Cited by 12 publications

(3 citation statements)

References 9 publications

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“…In Table 1 the results of fluorescence quenching studied obtained for time-resolved measurements are collected. Fluorescence lifetimes measured are in a reasonable agreement with literature data [27–31]. …”

Section: Resultssupporting

confidence: 90%

Analysis of Fluorescence Quenching of Coumarin Derivatives by 4-Hydroxy-TEMPO in Aqueous Solution

et al. 2013

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The fluorescence quenching of different coumarin derivatives (7-hydroxy-4-methylcoumarin, 5,7-dimethoxycoumarin, 7-amino-4-methyl-3-coumarinylacetic acid, 7-ethoxy-4-methylcoumarin, 7-methoxycoumarin, 7-hydroxycoumarin, 7-hydroxy-4-methyl-3-coumarinylacetic acid and 7-amino-4-methylcoumarin) by 4-hydroxy-TEMPO in aqueous solutions at the room temperature was studied with the use of UV–Vis absorption spectroscopy as well as a steady-state and time-resolved fluorescence spectroscopy. In order to understand the mechanism of quenching the absorption and fluorescence emission spectra of all coumarins along with fluorescence decays were recorded under the action of 4-hydroxy-TEMPO. The Stern-Volmer plots (both from time-averaged and time-resolved measurements) displayed no positive (upward) deviation from a linearity. The fluorescence quenching mechanism was found to be entirely dynamic, what was additionally confirmed by the registration of Stern-Volmer plots at different temperatures. The Stern-Volmer quenching constants and bimolecular quenching rate constants were obtained for all coumarins studied at the room temperature. The findings demonstrate the possibility of developing an analytical method for the quantitative determination of the free radicals’ scavenger, 4-hydroxy-TEMPO.

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Section: Resultssupporting

confidence: 90%

Analysis of Fluorescence Quenching of Coumarin Derivatives by 4-Hydroxy-TEMPO in Aqueous Solution

et al. 2013

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“…Usually a fluorophore, an enzyme, or a radioisotope marker can be used as the probe molecule (1,2). Furthermore, using time-resolved fluorescence measurements, the biological analytes also can be tagged with multiplex dyes, which show similar excitation and emission spectra characteristics but have different fluorescence lifetimes (3)(4)(5). Although these labels are specifically designed to receive a sensitive assay with the detection limit even down to the single molecule level, the chemical procedures involved in attachment of a label are time consuming, expensive, and labor intensive and may lead to additional losses of valuable sample material due to purification steps.…”

Section: Introductionmentioning

confidence: 99%

Autofluorescence Detection in Analytical Chemistry and Biochemistry

Seeger

2010

Applied Spectroscopy Reviews

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Label-free detection of molecules and particles based on native fluorescence excited at the ultraviolet (UV) region of the electromagnetic spectrum shows great potential for molecular analysis in life sciences. It offers simple, low-cost, and fast methods for sensitive detection of important biological analytes in nature forms. This article reviews the most significant research on label-free native fluorescence detection in analytical chemistry and biochemistry, with particular focus on the instrumental requirements and applications that cover papers published to the beginning of 2009. Future development directions for label-free optical biosensors based on native fluorescence are discussed.

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“…On the other hand, at concentrations of 10 -10 mol/L and above, most of the time more than one molecular complex is present in the detection volume. A cross correlation analysis using two different antibodies tagged with two dyes of different emission wavelengths or multiplex dyes of different fluorescence lifetimes − becomes difficult. Hence, the small dynamic range of affinity assays based on single-molecule detection in solution is not sufficient, and the lower detection limit is often too high for many applications.…”

mentioning

confidence: 99%

Counting of Single Protein Molecules at Interfaces and Application of This Technique in Early-Stage Diagnosis

et al. 1998

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The fluorescence-based detection and counting of single protein molecules after specific binding to antibodies at interfaces is presented. A diode laser was used as the excitation source. The unspecific binding at the interface has been reduced to a level of only 0.1% of the maximum signal level. At present, the detection limit of this molecule-counting process is in the range of 10(-17) mol/L, and the dynamic range of the signal corresponds to 7 orders of magnitude of antigen concentration, but these values are not limiting. As a preliminary application in early-stage diagnosis, we have investigated the detection of a single cardiac actin molecule in human plasma, which is of interest in myocardial infarction diagnosis.

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Simultaneous antigen detection using multiplex dyes

Cited by 12 publications

References 9 publications

Analysis of Fluorescence Quenching of Coumarin Derivatives by 4-Hydroxy-TEMPO in Aqueous Solution

Analysis of Fluorescence Quenching of Coumarin Derivatives by 4-Hydroxy-TEMPO in Aqueous Solution

Autofluorescence Detection in Analytical Chemistry and Biochemistry

Counting of Single Protein Molecules at Interfaces and Application of This Technique in Early-Stage Diagnosis

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