title: Photocleavable DNA Barcoding Antibodies for Multiplexed Protein Analysis in Single Cells.
abstract: We describe a DNA-barcoded antibody sensing technique for single cell protein analysis in which the barcodes are photocleaved and digitally detected without amplification steps (Ullal et al., Sci Transl Med 6:219, 2014). After photocleaving the unique ~70 mer DNA barcodes we use a fluorescent hybridization technology for detection, similar to what is commonly done for nucleic acid readouts. This protocol offers a simple method for multiplexed protein detection using 100+ antibodies and can be performed on clinical samples as well as single cells.
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, CPZN 5206, Boston, MA, 02114, USA.
authors.forename: Adeeti V
authors.initials: AV
authors.lastname: Ullal
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, CPZN 5206, Boston, MA, 02114, USA. rweissleder@mgh.harvard.edu.
authors.forename: Ralph
authors.initials: R
authors.lastname: Weissleder
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D058893:DNA Barcoding, Taxonomic / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry*; D008958:Models, Molecular; D009693:Nucleic Acid Hybridization / Q000379:methods; D010782:Photolysis; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*
chemical_list: D000906:Antibodies; D018796:Immunoconjugates; D011506:Proteins; D004247:DNA
keywords: Antibody conjugation; DNA barcoding; Fine needle aspirates; Multiplexing; Pathway analysis; Proteomics; Single cell analysis