Antti Vaheri

B Akerstrom

BH Nilson

D Klostermeier

EP Diamandis

H Bazin

Jussi Hepojoki

JW Stoop

K Blomberg

K Stenroos

Klaus Hedman

L Bjorck

M De Chateau

M Gabourdes

N Hildebrandt

O Gaillard

Olli Vapalahti

PA Maple

RP Ekins

S Saraheimo

Sabato D'Auria

Satu Hepojoki

T Förster

Visa Nurmi

Z Zheng

PLoS ONE

PubMed

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A Protein L -Based Immunodiagnostic Approach Utilizing Time-Resolved Förster Resonance Energy Transfer

Hepojoki, Satu

Nurmi, Visa

Vaheri, Antti

Hedman, Klaus

Vapalahti, Olli

Hepojoki, Jussi

Helsingin yliopiston digitaalinen arkisto

A Protein L -Based Immunodiagnostic Approach Utilizing Time-Resolved Forster Resonance Energy Transfer

Chelated lanthanides such as europium (Eu) have uniquely long fluorescence emission half-lives permitting their use in time-resolved fluorescence (TRF) assays. In Förster resonance energy transfer (FRET) a donor fluorophore transfers its emission energy to an acceptor fluorophore if in sufficiently close proximity. The use of time-resolved (TR) FRET minimizes the autofluorescence of molecules present in biological samples. In this report, we describe a homogenous immunoassay prototype utilizing TR-FRET for detection of antibodies in solution. The assay is based on labeled protein L, a bacterial protein that binds to immunoglobulin (Ig) light chain, and labeled antigen, which upon association with the same Ig molecule produce a TR-FRET active complex. We show that the approach is functional and can be utilized for both mono- and polyvalent antigens. We also compare the assay performance to that of another homogenous TR-FRET immunoassay reported earlier. This novel assay may have wide utility in infectious disease point-of-care diagnostics

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A protein L-based immunodiagnostic approach utilizing time-resolved Förster resonance energy transfer.

http://dx.doi.org/10.1371/journal.pone.0106432

A Protein L -Based Immunodiagnostic Approach Utilizing Time-Resolved Forster Resonance Energy Transfer

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Helsingin yliopiston digitaalinen arkisto

Directory of Open Access Journals