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Issue 9, 2010
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A homogeneous time-resolved fluoroimmunoassay (TR-FIA) using antibody mediated luminescence quenching

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Abstract

The determination of low-molecular weight substances (haptens) is demonstrated with a homogeneous time-resolved immunoassay using antibody-induced luminescence quenching. Our novel assay technology uses the newly developed monoclonal antibody (G24-BA9) to quench the luminescence of europium trisbipyridine (EuTBP). We performed a competitive biotin immunoassay including an EuTBPbiotin conjugate, the anti-EuTBP antibody G24-BA9 and streptavidin as assay components. Steric hindrance allows only the binding of either G24-BA9 (to the EuTBP moiety) or streptavidin (to the biotin moiety) to the EuTBPbiotin conjugate. Addition of the analyte biotin resulted in the binding of streptavidin to biotin and a concomitant preferred binding of G24-BA9 to EuTBPbiotin. Since G24-BA9 quenches the luminescence of EuTBP within the conjugate, the luminescence signal could be used to indicate and quantify the presence of free biotin in the system. All experiments were carried out in solution in the presence of 5% serum demonstrating the possibility of using our novel assay for a very fast determination of low molecular weight substances in biological fluids.

Graphical abstract: A homogeneous time-resolved fluoroimmunoassay (TR-FIA) using antibody mediated luminescence quenching

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Publication details

The article was received on 10 May 2010, accepted on 16 Jun 2010 and first published on 22 Jul 2010


Article type: Paper
DOI: 10.1039/C0AY00306A
Anal. Methods, 2010,2, 1298-1301

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    A homogeneous time-resolved fluoroimmunoassay (TR-FIA) using antibody mediated luminescence quenching

    F. Sellrie, M. Beck, N. Hildebrandt and B. Micheel, Anal. Methods, 2010, 2, 1298
    DOI: 10.1039/C0AY00306A

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