Detection of Alkynes via Click Chemistry with a Brominated Coumarin Azide by Simultaneous Fluorescence and Isotopic Signatures in Mass Spectrometry
Abstract
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.
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The practice of labeling functional groups in uncharacterized NP mixtures using a non-targeted metabolomics approach is uncommon in NP discovery, and there are currently only a few existing examples.
“Although these approaches have been in certain cases efficiently applied for NP discovery, the practice of labeling functional groups in uncharacterized NP mixtures using a non-targeted metabolomics approach is uncommon in NP discovery, and there are currently only a few existing examples.124–128”
From Connecting metabolome and phenotype: recent advances in functional metabolomics tools for the identification of bioactive natural products (2024) · cites this paper as [125]
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- Authors
- Yang, L., Chumsae, C., Kaplan, J. B., Moulton, K. R., Wang, D., Lee, D. H., & Zhou, Z. S.
- Affiliations
- AbbVie Bioresearch Center, 100 Research Drive, Worcester, Massachusetts 01605, United States
- Northeastern University (US)
- Corresponding author
- Chumsae, Chris
- Venue
- Bioconjugate Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2017-09-11
- Volume
- 28
- Issue
- 9
- Pages
- 2302-2309
- ISSN
- 1043-1802 (ISSN-L)
- 1520-4812
- OpenAlex
- W2746101075
- Cited by
- 27
- References
- 48
- Citations by year
- 2026: 2 · 2025: 1 · 2024: 7 · 2023: 2 · 2022: 1 · 2021: 2 · 2020: 4 · 2019: 6 · 2018: 1 · 2017: 1
- FWCI
- 1.37
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Bioconjugate Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of General Medical Sciences (1 R01 GM101396)
- Topics
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- click chemistry, 1,2,3-triazoles, fluorogenic labeling, isotopic signatures, monoclonal antibody conjugation, bioconjugation, fluorescence detection, mass spectrometry identification
- MeSH terms
- Alkynes, Amino Acid Sequence, Animals, Antibodies, Monoclonal, Azides, Coumarins, Cricetulus, Fluorescence, Fluorescent Dyes, Humans, Models, Molecular, Recombinant Proteins, Spectrometry, Fluorescence, Mass Spectrometry, Triazoles, CHO Cells, Halogenation, Click Chemistry
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