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Issue 4, 2011
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Blue fluorescent dye-protein complexes based on fluorogenic cyanine dyes and single chain antibody fragments

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Abstract

Fluoromodules are complexes formed upon the noncovalent binding of a fluorogenic dye to its cognate biomolecular partner, which significantly enhances the fluorescence quantum yield of the dye. Previously, several single-chain, variable fragment (scFv) antibodies were selected from a yeast cell surface-displayed library that activated fluorescence from a family of unsymmetrical cyanine dyes covering much of the visible and near-IR spectrum. The current work expands our repertoire of genetically encodable scFv-dye pairs by selecting and characterizing a group of scFvs that activate fluorogenic violet-absorbing, blue-fluorescing cyanine dyes, based on oxazole and thiazole heterocycles. The dye binds to both yeast cell surface-displayed and soluble scFvs with low nanomolar Kd values. These dye-protein fluoromodules exhibit high quantum yields, approaching unity for the brightest system. The promiscuity of these scFvs with other fluorogenic cyanine dyes was also examined. Fluorescence microscopy demonstrates that the yeast cell surface-displayed scFvs can be used for multicolor imaging. The prevalence of 405 nm lasers on confocal imaging and flow cytometry systems make these new reagents potentially valuable for cell biological studies.

Graphical abstract: Blue fluorescent dye-protein complexes based on fluorogenic cyanine dyes and single chain antibody fragments

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Publication details

The article was received on 18 Jul 2010, accepted on 18 Nov 2010 and first published on 22 Dec 2010


Article type: Paper
DOI: 10.1039/C0OB00444H
Citation: Org. Biomol. Chem., 2011,9, 1012-1020
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    Blue fluorescent dye-protein complexes based on fluorogenic cyanine dyes and single chain antibody fragments

    K. J. Zanotti, G. L. Silva, Y. Creeger, K. L. Robertson, A. S. Waggoner, P. B. Berget and B. A. Armitage, Org. Biomol. Chem., 2011, 9, 1012
    DOI: 10.1039/C0OB00444H

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