Issue 3, 2013

Obelin mutants as reporters in bioluminescent dual-analyte binding assay

Abstract

Two obelin mutants distinctly different in the color of bioluminescence were successfully applied to simultaneous detection of two analytes in a single well. The flash-type signals were triggered by single injection of Ca2+ and were discriminated by the spectral and time resolutions. The technique was developed in a high-throughput format, and applied to simultaneous immunoassay of two gonadotropic hormones ā€“ luteinizing (lutropin or hLH) and follicle stimulating (hFSH) ā€“ in clinical sera, and to simultaneous detection of two gene allelic variants at single nucleotide polymorphism genotyping of the human F5 gene encoding factor V Leiden polymorphism 1691 Gā†’A (R506Q). A considerable number of samples were investigated and the results obtained were in good correlation with those obtained by using traditional techniques.

Graphical abstract: Obelin mutants as reporters in bioluminescent dual-analyte binding assay

Article information

Article type
Paper
Submitted
03 Sep 2012
Accepted
28 Nov 2012
First published
29 Nov 2012

Anal. Methods, 2013,5, 636-640

Obelin mutants as reporters in bioluminescent dual-analyte binding assay

V. V. Krasitskaya, A. N. Kudryavtsev, O. Shimomura and L. A. Frank, Anal. Methods, 2013, 5, 636 DOI: 10.1039/C2AY25976A

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