Click chemistry for labeling and detection of biomolecules

Brian Agnew; Suzanne Buck; Tamara Nyberg; Jolene Bradford; Scott Clarke; Kyle Gee

doi:10.1117/12.762175

22 February 2008 Click chemistry for labeling and detection of biomolecules

Brian Agnew, Suzanne Buck, Tamara Nyberg, Jolene Bradford, Scott Clarke, Kyle Gee

Proceedings Volume 6867, Molecular Probes for Biomedical Applications II; 686708 (2008) https://doi.org/10.1117/12.762175
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

Described herein is an application of the copper(I)-catalyzed Huisgen [3+2] cycloaddition between azides and alkynes, or click chemistry, for the universal two-step detection of biological macromolecules. The first step involves the metabolic incorporation of an azide or alkyne probe into a macromolecule of interest. The second step involves the click chemistry conjugation of the labeled macromolecule with a partner alkyne or azide-reactive fluorescent probe to form a stable triazole ring conjugate. The fluorescently tagged molecules can be subsequently detected by a number of different fluorescent readout platforms including flow cytometry, fluorescence imaging, and 1-D/2-D gel imaging. We demonstrate application of this technology in two different labeling schemes. First, the labeling of newly synthesized DNA in a novel cell proliferation assay, and second, in the labeling of specific glycoprotein subclasses for biomarker discovery applications. In each case, azide or alkyne probes are introduced metabolically with subsequent detection using click chemistry. Utilization of the cellular enzymatic machinery for high-fidelity target molecule labeling combined with the superior efficiency of click chemistry detection results in highly versatile macromolecular labeling platforms that are unmatched in sensitivity and selectivity.

Citation Download Citation

Brian Agnew, Suzanne Buck, Tamara Nyberg, Jolene Bradford, Scott Clarke, and Kyle Gee "Click chemistry for labeling and detection of biomolecules", Proc. SPIE 6867, Molecular Probes for Biomedical Applications II, 686708 (22 February 2008); https://doi.org/10.1117/12.762175

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available