On-Chip Analysis of Protein Secretion from Single Cells Using Microbead Biosensors

The profiling of the effector functions of single immune cellsincluding cytokine secretioncan lead to a deeper understanding of how the immune system operates and to potential diagnostics and therapeutical applications. Here, we report a microfluidic device that pairs single cells and antibody-fun...

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Bibliographic Details
Published in:ACS sensors 2023-02, Vol.8 (2), p.655-664
Main Authors: Cedillo-Alcantar, Diana F., Rodriguez-Moncayo, Roberto, Maravillas-Montero, Jose L., Garcia-Cordero, Jose L.
Format: Article
Language:English
Subjects:
Antibodies
Biosensing Techniques
Cytokines
Humans
Interleukin-8
Microspheres
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Summary:The profiling of the effector functions of single immune cellsincluding cytokine secretioncan lead to a deeper understanding of how the immune system operates and to potential diagnostics and therapeutical applications. Here, we report a microfluidic device that pairs single cells and antibody-functionalized microbeads in hydrodynamic traps to quantitate cytokine secretion. The device contains 1008 microchambers, each with a volume of ∼500 pL, divided into six different sections individually addressed to deliver an equal number of chemical stimuli. Integrating microvalves allowed us to isolate cell/bead pairs, preventing cross-contamination with factors secreted by adjacent cells. We implemented a fluorescence sandwich immunoassay on the biosensing microbeads with a limit of detection of 9 pg/mL and were able to detect interleukin-8 (IL-8) secreted by single blood-derived human monocytes in response to different concentrations of LPS. Finally, our platform allowed us to observe a significant decrease in the number of IL-8-secreting monocytes when paracrine signaling becomes disrupted. Overall, our platform could have a variety of applications for which the analysis of cellular function heterogeneity is necessary, such as cancer research, antibody discovery, or rare cell screening.
ISSN:2379-3694
2379-3694
DOI:10.1021/acssensors.2c02148