Highly Multiplexed Single-Cell In Situ Protein Analysis with Cleavable Fluorescent Antibodies

Manas Mondal, Renjie Liao, Lu Xiao, Taylor Eno, Jia Guo

Research output: Contribution to journalArticlepeer-review

44 Scopus citations


Limitations on the number of proteins that can be quantified in single cells in situ impede advances in our deep understanding of normal cell physiology and disease pathogenesis. Herein, we present a highly multiplexed single-cell in situ protein analysis approach that is based on chemically cleavable fluorescent antibodies. In this method, antibodies tethered to fluorophores through a novel azide-based cleavable linker are utilized to detect their protein targets. After fluorescence imaging and data storage, the fluorophores coupled to the antibodies are efficiently cleaved without loss of protein target antigenicity. Upon continuous cycles of target recognition, fluorescence imaging, and fluorophore cleavage, this approach has the potential to quantify over 100 different proteins in individual cells at optical resolution. This single-cell in situ protein profiling technology will have wide applications in signaling network analysis, molecular diagnosis, and cellular targeted therapies.

Original languageEnglish (US)
Pages (from-to)2636-2639
Number of pages4
JournalAngewandte Chemie - International Edition
Issue number10
StatePublished - Mar 1 2017


  • fluorescent probes
  • immunoassays
  • protein expression
  • proteomics

ASJC Scopus subject areas

  • Catalysis
  • General Chemistry


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