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Updated: Jan 30, 2026

Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy
Published on: April 25, 2021
Bimodal Detection of Proteins by 129 Xe NMR and Fluorescence Spectroscopy
Emilie Mari1,2, Yasmina Bousmah2, Céline Boutin1
1NIMBE, CEA, CNRS, Université de Paris Saclay, CEA Saclay, 91191, Gif-sur-Yvette, France.
Abstract:
A full understanding of biological phenomena involves sensitive and noninvasive detection. Herein, we report the optimization of a probe for intracellular proteins that combines the advantages of fluorescence and hyperpolarized 129 Xe NMR spectroscopy detection. The fluorescence detection part is composed of six residues containing a tetracysteine tag (-CCXXCC-) genetically incorporated into the protein of interest and of a small organic molecule, CrAsH. CrAsH becomes fluorescent if it binds to the tetracysteine tag. The part of the biosensor that enables detection by means of 129 Xe NMR spectroscopy, which is linked to the CrAsH moiety by a spacer, is based on a cryptophane core that is fully suited to reversibly host xenon. Three different peptides, containing the tetracysteine tag and four organic biosensors of different stereochemistry, are benchmarked to propose the best couple that is fully suited for the in vitro detection of proteins.
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