Detection of Alkynes via Click Chemistry with a Brominated Coumarin Azide by Simultaneous Fluorescence and Isotopic Signatures in Mass Spectrometry

Alkynes are a key component of click chemistry and used for a wide variety of applications including bioconjugation, selective tagging of protein modifications, and labeling of metabolites and drug targets. However, challenges still exist for detecting alkynes because most 1,2,3-triazole products fr...

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Bibliographic Details
Published in:Bioconjugate chemistry 2017-09, Vol.28 (9), p.2302-2309
Main Authors: Yang, Lihua, Chumsae, Chris, Kaplan, Jenifer B, Moulton, Kevin Ryan, Wang, Dongdong, Lee, David H, Zhou, Zhaohui Sunny
Format: Article
Language:English
Subjects:
Alkynes
Alkynes - analysis
Amino Acid Sequence
Animals
Antibodies, Monoclonal - analysis
Azides - chemistry
Biomolecules
Bromination
Chemical compounds
Chemical synthesis
Chemistry
CHO Cells
Click Chemistry - methods
Coumarin
Coumarins - chemistry
Cricetulus
Fluorescence
Fluorescent Dyes - chemistry
Halogenation
Humans
Hydrocarbons
Mass spectrometry
Mass Spectrometry - methods
Mass spectroscopy
Metabolites
Models, Molecular
Monoclonal antibodies
Recombinant Proteins - analysis
Spectrometry, Fluorescence - methods
Spectroscopy
Triazoles - chemistry
Workflow
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Summary:Alkynes are a key component of click chemistry and used for a wide variety of applications including bioconjugation, selective tagging of protein modifications, and labeling of metabolites and drug targets. However, challenges still exist for detecting alkynes because most 1,2,3-triazole products from alkynes and azides do not possess distinct intrinsic properties that can be used for their facile detection by either fluorescence or mass spectrometry. To address this critical need, a novel brominated coumarin azide was used to tag alkynes and detect alkyne-conjugated biomolecules. This tag has several useful properties: first, it is fluorogenic and the click-chemistry products are highly fluorescent and quantifiable; second, its distinct isotopic pattern facilitates identification by mass spectrometry; and third, its click-chemistry products form a unique pair of reporter ions upon fragmentation that can be used for the quick screening of data. Using a monoclonal antibody conjugated with alkynes, a general workflow has been developed and examined comprehensively.
ISSN:1043-1802
1520-4812
DOI:10.1021/acs.bioconjchem.7b00354