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 1 2021
Accepted
18 3 2021
First published
20 3 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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