Immuno-Hybridization Chain Reaction for Enhancing Detection of Individual Cytokine-Secreting Human Peripheral Mononuclear Cells

We present here a new method to enhance the detection of secreted cytokines and chemokines from single human mononuclear cells. The technique uses a hybridization chain reaction (HCR) to amplify signals resulting from sandwich immunoassays. This immuno-HCR employs oligonucleotide-based initiators co...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2011-09, Vol.83 (17), p.6890-6895
Main Authors: Choi, Jonghoon, Routenberg Love, Kerry, Gong, Yuan, Gierahn, Todd M, Love, J. Christopher
Format: Article
Language:English
Subjects:
Analytical chemistry
Antibodies, Immobilized - immunology
Cells
Cells, Cultured
Chemistry
Chemokines - analysis
Chemokines - immunology
Cytokines
Cytokines - analysis
Cytokines - immunology
Dimethylpolysiloxanes - chemistry
Exact sciences and technology
Fluorescence
Fluorescent Dyes - chemistry
Glass - chemistry
Humans
Hybridization
Immunoassay
Immunoassay - methods
Leukocytes, Mononuclear - immunology
Leukocytes, Mononuclear - metabolism
Miscellaneous
Oligonucleotides - chemistry
Protein Array Analysis - methods
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Summary:We present here a new method to enhance the detection of secreted cytokines and chemokines from single human mononuclear cells. The technique uses a hybridization chain reaction (HCR) to amplify signals resulting from sandwich immunoassays. This immuno-HCR employs oligonucleotide-based initiators covalently linked to antibodies to propagate a chain reaction of hybridization events involving a pair of complementary hairpin oligomers bearing fluorescent labels. Integrating this strategy for signal amplification with microengraving (a soft lithographic method for printing arrays of secreted proteins from thousands of single cells) improves both the limits of detection and sensitivity for cytokines and chemokines captured from individual cells by an average of 200-fold relative to methods for direct detection by fluoresence. This approach should enhance the utility of microengraving for defining the immunological signatures of diseases and responses to interventional therapies based on multiplexed single-cell analysis.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac2013916