High-throughput secretomic analysis of single cells to assess functional cellular heterogeneity

Yao Lu, Jonathan J. Chen, Luye Mu, Qiong Xue, Yu Wu, Pei Hsun Wu, Jie Li, Alexander O. Vortmeyer, Kathryn Miller-Jensen, Denis Wirtz, Rong Fan

Research output: Contribution to journalArticle

90 Scopus citations

Abstract

Secreted proteins dictate a range of cellular functions in human health and disease. Because of the high degree of cellular heterogeneity and, more importantly, polyfunctionality of individual cells, there is an unmet need to simultaneously measure an array of proteins from single cells and to rapidly assay a large number of single cells (more than 1000) in parallel. We describe a simple bioanalytical assay platform consisting of a large array of subnanoliter microchambers integrated with high-density antibody barcode microarrays for highly multiplexed protein detection from over a thousand single cells in parallel. This platform has been tested for both cell lines and complex biological samples such as primary cells from patients. We observed distinct heterogeneity among the single cell secretomic signatures that, for the first time, can be directly correlated to the cells' physical behavior such as migration. Compared to the state-of-the-art protein secretion assay such as ELISpot and emerging microtechnology-enabled assays, our approach offers both high throughput and high multiplicity. It also has a number of clinician-friendly features such as ease of operation, low sample consumption, and standardized data analysis, representing a potentially transformative tool for informative monitoring of cellular function and immunity in patients.

Original languageEnglish (US)
Pages (from-to)2548-2556
Number of pages9
JournalAnalytical Chemistry
Volume85
Issue number4
DOIs
StatePublished - Feb 19 2013

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ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Lu, Y., Chen, J. J., Mu, L., Xue, Q., Wu, Y., Wu, P. H., Li, J., Vortmeyer, A. O., Miller-Jensen, K., Wirtz, D., & Fan, R. (2013). High-throughput secretomic analysis of single cells to assess functional cellular heterogeneity. Analytical Chemistry, 85(4), 2548-2556. https://doi.org/10.1021/ac400082e