Immuno-Hybridization Chain Reaction for Enhancing Detection of Individual Cytokine-Secreting Human Peripheral Mononuclear Cells
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nihms317360.pdf
Description
Accepted version
Size
1.49 MB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Choi, Jonghoon
Routenberg Love, Kerry
Gong, Yuan
Gierahn, Todd M
Love, J Christopher
Date Issued
2011
Journal
Analytical Chemistry
Publisher
American Chemical Society (ACS)
Citation
Choi, Jonghoon, et al. "Immuno-Hybridization Chain Reaction for Enhancing Detection of Individual Cytokine-Secreting Human Peripheral Mononuclear Cells." Analytical Chemistry 83 17 (2011): 6890-95.
Version
Author's final manuscript
Abstract
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. © 2011 American Chemical Society.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
Ragon Institute of MGH, MIT and Harvard
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1021/ac2013916
