Membrane-based lateral flow assay devices that utilize phosphorescent detection

a membrane-based assay and detection method technology, applied in biomass after-treatment, instruments, enzymology, etc., can solve the problems of limiting the use of phosphorescent detection in most practical assay applications, many of the proposed techniques are simply ill equipped for lateral flow,

Inactive Publication Date: 2005-05-26
KIMBERLY-CLARK WORLDWIDE INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, many of the proposed techniques fail to solve the problem of quenching.
Specifically, oxygen and water are strong quenchers of triplet states and may cause decay of the phosphorescence signal, thereby limiting its use in most practical assay applications.
In addition, many of the techniques that have been proposed are simply ill equipped for use in lateral flow, membrane-based devices.
However, background interference becomes increasingly problematic at such low analyte concentrations because the phosphorescent intensity is relatively low.
This fluorescence can interfere with the accuracy of phosphorescence measurements.

Method used

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  • Membrane-based lateral flow assay devices that utilize phosphorescent detection
  • Membrane-based lateral flow assay devices that utilize phosphorescent detection
  • Membrane-based lateral flow assay devices that utilize phosphorescent detection

Examples

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example 1

[0063] The ability to encapsulate platinum(II)tetra-meso-fluorophenylporphine (Pt(II)-MTPFP) in carboxylated latex particles was demonstrated. Initially, 8 milligrams of a carboxylated latex particle suspension (0.3-micrometer particle size, available from Bangs Laboratories, Inc) was washed twice with ethanol and then suspended in 100 microliters of ethanol. 100 micrograms of Pt(II)MTPFP in 55 microliters of methylene chloride and 45 microliters of ethanol was added to the particle suspension. The mixture was gently shaken for 20 minutes. Then, 100 microliters of water was added. The mixture was then shaken overnight. Approximately 50 vol. % of the solvent was then removed by air stream. 1 milliliter of ethanol was added and centrifuged, after which the supernatant was discarded. The particles were further washed twice with ethanol and then twice with water. The washed particles were suspended by bath sonication in 1.5 milliliter of water for storage. The particles exhibited very s...

example 2

[0064] The encapsulation of platinum(II)tetra-meso-fluorophenylporphine in polyacrylonitrile particles was demonstrated. 118.5 milligrams of polyacrylonitrile and 1.19 milligrams of platinum(II)tetra-meso-fluorophenylporphine were dissolved in 25 milliliters of DMF. 125 milliliters of water was added to the mixture under vigorous stirring. Thereafter, 1 milliliter of a saturated sodium chloride aqueous solution was added to precipitate the dispersed particles. The mixture was centrifuged for 15 minutes and then washed twice with a sodium chloride solution (5 wt. %) and three times with 50 milliliters of water. The residue was then heated for 15 minutes at 70° C. and centrifuged in a phosphate buffer. This procedure is substantially identical to that described in Bioconjugated Chemistry, Kurner, 12, 883-89 (2001), with the exception that the ruthenium complexes were replaced with platinum(II)tetra-meso-fluorophenylporphine.

[0065] The resulting encapsulated particles exhibited very s...

example 3

[0066] The ability to conjugate an antibody to phosphorescent particles was demonstrated. The particles were carboxylated latex phosphorescent particles having a particle size of 0.20 micrometers, 0.5% solids, and exhibiting phosphorescence at an emission wavelength of 650 nanometers when excited at a wavelength of 390 nanometers. The particles were obtained from Molecular Probes, Inc. under the name “FluoSpheres.”

[0067] Initially, 500 microliters of the particles were washed once with 1 milliliter of a carbonate buffer and twice with 2-[N-morpholino]ethanesulfonic acid (MES) buffer (pH: 6.1, 20 millimolar) using a centrifuge. The washed particles were re-suspended in 250 microliters of MES. Thereafter, 3 milligrams of carbodiimide (Polysciences, Inc.) was dissolved in 250 microliters of MES and added to the suspended particles. The mixture was allowed to react at room temperature for 30 minutes on a shaker. The activated particles were then washed twice with a borate buffer (Polysc...

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Abstract

A lateral flow, membrane-based assay device for detecting the presence or quantity of an analyte residing in a test sample is provided. The device utilizes phosphorescence to detect the signals generated by excited phosphorescent labels. The labels may have a long emission lifetime so that background interference from many sources, such as scattered light and autofluorescence, is practically eliminated during detection. In addition, the phosphorescent labels may be encapsulated within particles to shield the labels from quenchers, such as oxygen or water, which might disrupt the phosphorescent signal.

Description

BACKGROUND OF THE INVENTION [0001] Phosphorescence is the result of a three-stage process. In the first stage, energy is supplied by an external source, such as an incandescent lamp or a laser, and absorbed by the phosphorescent compound, creating excited electronic triplet states (as opposed to fluorescence, which only has a singlet excited state). In the second stage, the excited states exist for a finite time during which the phosphorescent compound undergoes conformational changes and is also subject to a multitude of possible interactions with its molecular environment. During this time, the energy of the excited states is partially dissipated, yielding relaxed states from which phosphorescence emission originates. The third stage is the phosphorescence emission stage wherein energy is emitted, returning the phosphorescence compound to its ground states. The emitted energy is lower than its excitation energy (light or laser) and thus of a longer wavelength. This shift or differ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07D487/22C12M1/34G01N33/53G01N33/537G01N33/543G01N33/558G01N33/58
CPCG01N33/54393G01N33/582G01N33/587G01N21/6428G01N33/5302Y10S436/81Y10S435/97Y10S435/81Y10S435/805G01N33/558G01N33/54313Y10S436/805
Inventor SONG, XUEDONG
Owner KIMBERLY-CLARK WORLDWIDE INC
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