2. Academic Validation
  3. Click-based amplification: designed to facilitate various target labelling modes with ultralow background amplification

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

  • RSC Chem Biol. 2021 Mar 20;2(3):906-916. doi: 10.1039/d1cb00002k.
Jinyi Bai 1 2,  Fusheng Guo 1 2,  Mengyao Li 3 4 5,  Yulong Li 2 3 4 5,  Xiaoguang Lei 1 2
Affiliations

Affiliations

  • 1 Beijing National Laboratory for Molecular Sciences, State Key Laboratory of Natural and Biomimetic Drugs, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Department of Chemical Biology, College of Chemistry and Molecular Engineering, Synthetic and Functional Biomolecules Center, Peking University Beijing 100871 People's Republic of China [email protected].
  • 2 Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University Beijing 100871 People's Republic of China [email protected].
  • 3 State Key Laboratory of Membrane Biology, Peking University School of Life Sciences Beijing China.
  • 4 PKU-IDG/McGovern Institute for Brain Research Beijing China.
  • 5 Chinese Institute for Brain Research Beijing China.
PMID: 34458817 DOI: 10.1039/d1cb00002k
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.

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