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A Modular, DNA‐Based Beacon for Single‐Step Fluorescence Detection of Antibodies and Other Proteins
Author(s) -
Ranallo Simona,
Rossetti Marianna,
Plaxco Kevin W.,
ValléeBélisle Alexis,
Ricci Francesco
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201505179
Subject(s) - chemistry , fluorophore , molecular beacon , nucleic acid , fluorescence , dna , biophysics , biochemistry , combinatorial chemistry , biology , physics , oligonucleotide , quantum mechanics
A versatile platform for the one‐step fluorescence detection of both monovalent and multivalent proteins has been developed. This system is based on a conformation‐switching stem–loop DNA scaffold that presents a small‐molecule, polypeptide, or nucleic‐acid recognition element on each of its two stem strands. The steric strain associated with the binding of one (multivalent) or two (monovalent) target molecules to these elements opens the stem, enhancing the emission of an attached fluorophore/quencher pair. The sensors respond rapidly (<10 min) and selectively, enabling the facile detection of specific proteins even in complex samples, such as blood serum. The versatility of the platform was demonstrated by detecting five bivalent proteins (four antibodies and the chemokine platelet‐derived growth factor) and two monovalent proteins (a Fab fragment and the transcription factor TBP) with low nanomolar detection limits and no detectable cross‐reactivity.