title: Highly Multiplexed Single-Cell In Situ Protein Analysis with Cleavable Fluorescent Antibodies.
abstract: 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.
authors.affiliation: Biodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, Arizona, 85287, USA.
authors.forename: Manas
authors.initials: M
authors.lastname: Mondal
authors.affiliation: Biodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, Arizona, 85287, USA.
authors.forename: Renjie
authors.initials: R
authors.lastname: Liao
authors.affiliation: Biodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, Arizona, 85287, USA.
authors.forename: Lu
authors.initials: L
authors.lastname: Xiao
authors.affiliation: Biodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, Arizona, 85287, USA.
authors.forename: Taylor
authors.initials: T
authors.lastname: Eno
authors.affiliation: Biodesign Institute & School of Molecular Sciences, Arizona State University, Tempe, Arizona, 85287, USA.
authors.forename: Jia
authors.initials: J
authors.lastname: Guo
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry*; D001386:Azides / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D015394:Molecular Structure; D061848:Optical Imaging; D011506:Proteins / Q000032:analysis*; D059010:Single-Cell Analysis
chemical_list: D000900:Anti-Bacterial Agents; D001386:Azides; D005456:Fluorescent Dyes; D011506:Proteins
keywords: fluorescent probes; immunoassays; protein expression; proteomics