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Clinical Application of Time-Resolved Fluorometric Assays

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Lanthanide Luminescence

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 7))

Abstract

The development of time-resolved fluoroimmunoassays (TRFIAs) was driven by the need to eliminate the use of radioactive isotopes in immunoassays and to develop automatic and ultrasensitive immunoassays. The use of time-resolved fluorescence became possible through the development of chelates that bound lanthanides with high affinity and could be coupled to proteins. This was combined with methods to dissociate the primary chelate together with the formation of a highly fluorescent new complex. The advantages of this technology are most efficiently used in sandwich assays using two antibodies, a solid phase antibody capturing the antigen and a detector antibody labeled with a lanthanide complex. This method facilitated a 10–100-fold lower detection limit in combination with an assay range covering 5–6 orders of magnitude. These advantages have facilitated elucidation of new biological phenomena and development of unique clinical applications. These have proved to be especially valuable for the determination of protein hormones and tumor markers. This review describes the clinical application of TRFIA.

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Abbreviations

ACT:

α1-Antichymotrypsin

Aio!:

All-in-one

BPH:

Benign prostatic hyperplasia

cpm:

Counts per minute

DELFIAs:

Dissociation-enhanced lanthanide fluoroimmunoassays

Dy:

Dysprosium

Eu:

Europium

fPSA:

Free PSA

FSH:

Follicle-stimulating hormone

GH:

Growth hormone

hCG:

Human chorionic gonadotropin

hCGβ:

β Subunit of human chorionic gonadotropin

hK2:

Human kallikrein 2

IEMA:

Immuno-enzymometric assays

IFMA:

Immunofluorometric assay

Ig:

Immunoglobulin

IRMA:

Immunoradiometric assay

KLK:

Kallikrein-related peptidase

LH:

Luteinizing hormone

MAb:

Monoclonal antibody

PCR:

Polymerase chain reaction

PSA:

Prostate-specific antigen

PSA-ACT:

Complex between PSA and ACT

RIA:

Radioimmunoassay

Sm:

Samarium

Tb:

Terbium

tPSA:

Total PSA (sum of free and complexed PSA)

TRFIA:

Time-resolved fluoroimmunoassay, time-resolved fluorometric assay

TR-FRET:

Time-resolved fluorescence resonance energy transfer

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  1. Department of Clinical Chemistry, Biomedicum, Helsinki University Central Hospital, Helsinki University, 00014, Helsinki, Finland

    Ulf-Håkan Stenman

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Correspondence to Ulf-Håkan Stenman .

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  1. , Department of Cell Biology and Anatomy, University of Turki, Tykistökatu 6A, Turku, 20520, Finland

    Pekka Hänninen

  2. , Department of Cell Biology and Anatomy, University of Turku, Tykistökatu 6A, Turku, 20520, Finland

    Harri Härmä

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Stenman, UH. (2010). Clinical Application of Time-Resolved Fluorometric Assays. In: Hänninen, P., Härmä, H. (eds) Lanthanide Luminescence. Springer Series on Fluorescence, vol 7. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2010_12

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