Thin solid europium(III) dye layers as donors in time-resolved fluorescence resonance energy transfer assays

Harri Härmä (Corresponding Author), Riikka Suhonen, Terho Kololuoma, Ari Kärkkäinen, Mika Hara, Pekka Hänninen

Research output: Contribution to journalArticleScientificpeer-review

6 Citations (Scopus)

Abstract

Lanthanide chelates and lanthanide nanoparticle labels are attractive donors for separation-free time-resolved fluorescence resonance energy transfer (TR-FRET) assays. In fully dyed nanoparticles, the inner volume of nanoparticle labels in TR-FRET assays are incapable of participating to energy transfer due to large distances to acceptors on the surface. Our interest was to study surface-based TR-FRET and, therefore, various europium(III) (Eu) chelate layers were investigated for TR-FRET efficiency. Eu(III) chelates incorporated in a siloxane layer, Eu(III) chelate covalently coupled on silanized surface and Eu(III) labeled protein surface were prepared and compared to nanoparticle-based TR-FRET. Energy transfer between the solid-phase donors and Cy5-labeled protein were obtained with signal-to-background ratios ranging from 1.2 to 9.9. In this study, a thin layer prepared using Eu(III)-labeled protein gave the most efficient TR-FRET. This thin donor layer was tested in a competitive separation-free immunoassay of human albumin (hAlb). hAlb was measured in a clinically relevant concentrations from 0.05 to 10 mg l−1 with the coefficient of variation ranging from 1.0% to 12.4%.

Original languageEnglish
Pages (from-to)6529-6534
Number of pages6
JournalApplied Surface Science
Volume255
Issue number13-14
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fingerprint

Europium
Assays
Coloring Agents
Dyes
Nanoparticles
Lanthanoid Series Elements
Rare earth elements
Proteins
Energy transfer
Labels
Albumins
Siloxanes
Fluorescence Resonance Energy Transfer
Membrane Proteins

Keywords

  • TR-FRET
  • europium
  • thin solid surface
  • immunoassay

Cite this

Härmä, Harri ; Suhonen, Riikka ; Kololuoma, Terho ; Kärkkäinen, Ari ; Hara, Mika ; Hänninen, Pekka. / Thin solid europium(III) dye layers as donors in time-resolved fluorescence resonance energy transfer assays. In: Applied Surface Science. 2009 ; Vol. 255, No. 13-14. pp. 6529-6534.
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abstract = "Lanthanide chelates and lanthanide nanoparticle labels are attractive donors for separation-free time-resolved fluorescence resonance energy transfer (TR-FRET) assays. In fully dyed nanoparticles, the inner volume of nanoparticle labels in TR-FRET assays are incapable of participating to energy transfer due to large distances to acceptors on the surface. Our interest was to study surface-based TR-FRET and, therefore, various europium(III) (Eu) chelate layers were investigated for TR-FRET efficiency. Eu(III) chelates incorporated in a siloxane layer, Eu(III) chelate covalently coupled on silanized surface and Eu(III) labeled protein surface were prepared and compared to nanoparticle-based TR-FRET. Energy transfer between the solid-phase donors and Cy5-labeled protein were obtained with signal-to-background ratios ranging from 1.2 to 9.9. In this study, a thin layer prepared using Eu(III)-labeled protein gave the most efficient TR-FRET. This thin donor layer was tested in a competitive separation-free immunoassay of human albumin (hAlb). hAlb was measured in a clinically relevant concentrations from 0.05 to 10 mg l−1 with the coefficient of variation ranging from 1.0{\%} to 12.4{\%}.",
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Thin solid europium(III) dye layers as donors in time-resolved fluorescence resonance energy transfer assays. / Härmä, Harri (Corresponding Author); Suhonen, Riikka; Kololuoma, Terho; Kärkkäinen, Ari; Hara, Mika; Hänninen, Pekka.

In: Applied Surface Science, Vol. 255, No. 13-14, 2009, p. 6529-6534.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Thin solid europium(III) dye layers as donors in time-resolved fluorescence resonance energy transfer assays

AU - Härmä, Harri

AU - Suhonen, Riikka

AU - Kololuoma, Terho

AU - Kärkkäinen, Ari

AU - Hara, Mika

AU - Hänninen, Pekka

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N2 - Lanthanide chelates and lanthanide nanoparticle labels are attractive donors for separation-free time-resolved fluorescence resonance energy transfer (TR-FRET) assays. In fully dyed nanoparticles, the inner volume of nanoparticle labels in TR-FRET assays are incapable of participating to energy transfer due to large distances to acceptors on the surface. Our interest was to study surface-based TR-FRET and, therefore, various europium(III) (Eu) chelate layers were investigated for TR-FRET efficiency. Eu(III) chelates incorporated in a siloxane layer, Eu(III) chelate covalently coupled on silanized surface and Eu(III) labeled protein surface were prepared and compared to nanoparticle-based TR-FRET. Energy transfer between the solid-phase donors and Cy5-labeled protein were obtained with signal-to-background ratios ranging from 1.2 to 9.9. In this study, a thin layer prepared using Eu(III)-labeled protein gave the most efficient TR-FRET. This thin donor layer was tested in a competitive separation-free immunoassay of human albumin (hAlb). hAlb was measured in a clinically relevant concentrations from 0.05 to 10 mg l−1 with the coefficient of variation ranging from 1.0% to 12.4%.

AB - Lanthanide chelates and lanthanide nanoparticle labels are attractive donors for separation-free time-resolved fluorescence resonance energy transfer (TR-FRET) assays. In fully dyed nanoparticles, the inner volume of nanoparticle labels in TR-FRET assays are incapable of participating to energy transfer due to large distances to acceptors on the surface. Our interest was to study surface-based TR-FRET and, therefore, various europium(III) (Eu) chelate layers were investigated for TR-FRET efficiency. Eu(III) chelates incorporated in a siloxane layer, Eu(III) chelate covalently coupled on silanized surface and Eu(III) labeled protein surface were prepared and compared to nanoparticle-based TR-FRET. Energy transfer between the solid-phase donors and Cy5-labeled protein were obtained with signal-to-background ratios ranging from 1.2 to 9.9. In this study, a thin layer prepared using Eu(III)-labeled protein gave the most efficient TR-FRET. This thin donor layer was tested in a competitive separation-free immunoassay of human albumin (hAlb). hAlb was measured in a clinically relevant concentrations from 0.05 to 10 mg l−1 with the coefficient of variation ranging from 1.0% to 12.4%.

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