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Traceless Cleavage of Protein–Biotin Conjugates under Biologically Compatible Conditions
Author(s) -
Cowell Joseph,
Buck Matthew,
Essa Ali H.,
Clarke Rebecca,
Vollmer Waldemar,
Vollmer Daniela,
Hilkens Catharien M.,
Isaacs John D.,
Hall Michael J.,
Gray Joe
Publication year - 2017
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201700214
Subject(s) - biotinylation , chemistry , avidin , biotin , biomolecule , linker , conjugate , fluorophore , bioorthogonal chemistry , combinatorial chemistry , streptavidin , chromatography , pretargeting , biochemistry , fluorescence , click chemistry , mathematical analysis , physics , mathematics , operating system , antibody , radioimmunotherapy , immunology , monoclonal antibody , biology , quantum mechanics , computer science
Biotinylation of amines is widely used to conjugate biomolecules, but either the resulting label is non‐removable or its removal leaves a tag on the molecule of interest, thus affecting downstream processes. We present here a set of reagents (RevAmines) that allow traceless, reversible biotinylation under biologically compatible, mild conditions. Release following avidin‐based capture is achieved through the cleavage of a (2‐(alkylsulfonyl)ethyl) carbamate linker under mild conditions (200 m m ammonium bicarbonate, pH 8, 16–24 h, room temperature) that regenerates the unmodified amine. The capture and release of biotinylated proteins and peptides from neutravidin, fluorescent labelling through reversible biotinylation at the cell surface and the selective enrichment of proteins from bacterial periplasm are demonstrated. The tags are easily prepared, stable and offer the potential for future application in proteomics, activity‐based protein profiling, affinity chromatography and bio‐molecule tagging and purification.