Issue 3, 2021

Click-based amplification: designed to facilitate various target labelling modes with ultralow background amplification

Abstract

We here describe a fluorescent signal amplification method termed “Click-based amplification” that can be well integrated with various click-labelling modes, including chemical labelling, genetic incorporation and covalent inhibitor probe mediated target labelling. Picolyl azide (pAz) was used as a functional group of a streptavidin-based amplifier to enhance the efficiency of click chemistry. Click-based amplification provided 3.0–12.7 fold amplification on fixed HeLa cells with different click-labelling modes. Click-based amplification has proven to be superior to tyramide signal amplification (TSA) in view of its low nonspecific amplification and high signal-to-noise ratio. Moreover, in terms of the challenging signal amplification of tissue specimens, Click-based amplification successfully achieved remarkable fluorescence enhancement on intestinal tissue slices of afatinib-N3 treated mice, which provided direct evidence of the presence of afatinib-N3 in the intestinal tissues and helped in revealing the off-target toxicity of afatinib. Collectively, these results illustrate that Click-based amplification could serve as a promising method for bioimaging studies.

Graphical abstract: Click-based amplification: designed to facilitate various target labelling modes with ultralow background amplification

Supplementary files

Article information

Article type
Paper
Submitted
06 Janv. 2021
Accepted
18 Marts 2021
First published
20 Marts 2021
This article is Open Access
Creative Commons BY license

RSC Chem. Biol., 2021,2, 906-916

Click-based amplification: designed to facilitate various target labelling modes with ultralow background amplification

J. Bai, F. Guo, M. Li, Y. Li and X. Lei, RSC Chem. Biol., 2021, 2, 906 DOI: 10.1039/D1CB00002K

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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