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Functional TCR T cell screening using single-cell droplet microfluidics

Segaliny, Aude I. and Li, Guideng and Kong, Lingshun and Ren, Ci and Chen, Xiaoming and Wang, Jessica K. and Baltimore, David and Wu, Guikai and Zhao, Weian (2018) Functional TCR T cell screening using single-cell droplet microfluidics. Lab on a Chip, 18 (24). pp. 3733-3749. ISSN 1473-0197. PMCID PMC6279597. doi:10.1039/c8lc00818c.

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Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to conduct fast and multiplexed functional analyses followed by recovery of the clone of interest. Here, we use droplet microfluidics for functional screening and real-time monitoring of single TCR T cell activation upon recognition of target tumor cells. Notably, our platform includes a tracking system for each clone as well as a sorting procedure with 100% specificity validated by downstream single cell reverse-transcription PCR and sequencing of TCR chains. Our TCR screening prototype will facilitate immunotherapeutic screening and development of T cell therapies.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Segaliny, Aude I.0000-0001-9670-8350
Li, Guideng0000-0003-0840-7262
Wang, Jessica K.0000-0003-1421-4969
Baltimore, David0000-0001-8723-8190
Zhao, Weian0000-0002-7794-9914
Additional Information:© 2018 The Royal Society of Chemistry. The article was received on 07 Aug 2018, accepted on 26 Oct 2018 and first published on 06 Nov 2018. This article is part of the themed collections: Lab on a Chip Emerging Investigators and Droplet-based single-cell sequencing. The authors would like to thank Chih Chun Yu for his help in finishing the TCR T cell activation experiments in droplets and assistance in droplet sorting, and Stephanie Wong for her help in single cell PCR work. The authors are also grateful to Cassandra Kiyomi Davis for her support in culturing the cells and performing cell viability test from droplets. This work is supported by the NIH (1DP2CA195763 and R21CA219225), and Amberstone Biosciences LLC: No. AB-208317. Author contributions: Conceptualization, A. I. S., G. L., G. W. and W. Z.; methodology, A. I. S., G. L., L. K., C. R., X. C. and G. W.; investigation, A. I. S, G. L., L. K., C. R., and J. K. W.; analysis, A. I. S.; writing: original draft, A. I. S. and W. Z.; editing, A. I. S., G. L., L. K., X. C., J. K. W., D. B., G. W. and W. Z.; supervision, A. I. S., D. B., G. W. and W. Z. Conflicts of interest: Weian Zhao is a co-founder of Amberstone Biosciences LLC that develops immunotherapeutics.
Funding AgencyGrant Number
Amberstone Biosciences LLCAB-208317
Issue or Number:24
PubMed Central ID:PMC6279597
Record Number:CaltechAUTHORS:20181106-104559775
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:90667
Deposited By: Tony Diaz
Deposited On:06 Nov 2018 18:56
Last Modified:16 Feb 2022 23:20

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