bc7b00354_si_001.pdf (1.65 MB)
Detection of Alkynes via Click Chemistry with a Brominated Coumarin Azide by Simultaneous Fluorescence and Isotopic Signatures in Mass Spectrometry
journal contribution
posted on 2017-08-21, 00:00 authored by Lihua Yang, Chris Chumsae, Jenifer B. Kaplan, Kevin Ryan Moulton, Dongdong Wang, David H. Lee, Zhaohui Sunny ZhouAlkynes
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.