A microfluidic-SERSplatform for isolation and immuno-phenotyping of antigen specific T-cells

Shuvashis Dey, Ramanathan Vaidyanathan, K. Kamil Reza, Jing Wang, Yuling Wang, Hendrik J. Nel, Soi Cheng Law, Jennifer Tyler, Jamie Rossjohn, Hugh H. Reid, Abu Ali Ibn Sina, Ranjeny Thomas, Matt Trau

Research output: Contribution to journalArticleResearchpeer-review

Abstract

T-cells play a major role in host defense mechanisms against many diseases. With the current growth of immunotherapy approaches, there is a strong need for advanced technologies to detect and characterize these immune cells. Herein, we present a simple approach for the isolation of antigen specific T-cells from the complex biological sample based on T-cell receptor (TCR) and peptide major histocompatibility complex (pMHC) interaction. Subsequently, we characterize those antigen specific T-cells by profiling TCR expression heterogeneity. Our approach utilizes an alternating current electrohydrodynamic (ac-EHD) based microfluidic platform for isolation and surface enhanced Raman scattering (SERS) for TCR expression profiling. The use of ac-EHD enables specific isolation of T-cells by generating a nanoscopic shear force at the double layer of the sensing surface which enhances the frequency of pMHC and TCR interactions and consequently shears off the nonspecific targets. TCR expression profiling of the isolated T-cells was performed by encoding them with SERS-labelled pMHCs followed by SERS detection in bulk as well as in single T-Cell. In proof-of-concept experiments, 56.93 ± 7.31% of the total CD4+T-cells were captured from an excess amount of nonspecific cells (e.g., PBMCs) with high specificity and sensitivity (0.005%). Moreover, TCR analysis data using SERS shows the heterogeneity in the T-cell receptor expression which can inform on the activation status of T-cells and the patient's response to immunotherapy. We believe that this approach may hold potential for numerous applications towards monitoring immune status, understanding therapeutic responses,and effective vaccine development.

Original languageEnglish
Pages (from-to)281-288
Number of pages8
JournalSensors and Actuators B: Chemical
Volume284
DOIs
Publication statusPublished - 1 Apr 2019

Keywords

  • Microfluidics
  • pMHC
  • SERS
  • T-cell
  • TCR

Cite this

Dey, S., Vaidyanathan, R., Reza, K. K., Wang, J., Wang, Y., Nel, H. J., ... Trau, M. (2019). A microfluidic-SERSplatform for isolation and immuno-phenotyping of antigen specific T-cells. Sensors and Actuators B: Chemical, 284, 281-288. https://doi.org/10.1016/j.snb.2018.12.099
Dey, Shuvashis ; Vaidyanathan, Ramanathan ; Reza, K. Kamil ; Wang, Jing ; Wang, Yuling ; Nel, Hendrik J. ; Law, Soi Cheng ; Tyler, Jennifer ; Rossjohn, Jamie ; Reid, Hugh H. ; Ibn Sina, Abu Ali ; Thomas, Ranjeny ; Trau, Matt. / A microfluidic-SERSplatform for isolation and immuno-phenotyping of antigen specific T-cells. In: Sensors and Actuators B: Chemical. 2019 ; Vol. 284. pp. 281-288.
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abstract = "T-cells play a major role in host defense mechanisms against many diseases. With the current growth of immunotherapy approaches, there is a strong need for advanced technologies to detect and characterize these immune cells. Herein, we present a simple approach for the isolation of antigen specific T-cells from the complex biological sample based on T-cell receptor (TCR) and peptide major histocompatibility complex (pMHC) interaction. Subsequently, we characterize those antigen specific T-cells by profiling TCR expression heterogeneity. Our approach utilizes an alternating current electrohydrodynamic (ac-EHD) based microfluidic platform for isolation and surface enhanced Raman scattering (SERS) for TCR expression profiling. The use of ac-EHD enables specific isolation of T-cells by generating a nanoscopic shear force at the double layer of the sensing surface which enhances the frequency of pMHC and TCR interactions and consequently shears off the nonspecific targets. TCR expression profiling of the isolated T-cells was performed by encoding them with SERS-labelled pMHCs followed by SERS detection in bulk as well as in single T-Cell. In proof-of-concept experiments, 56.93 ± 7.31{\%} of the total CD4+T-cells were captured from an excess amount of nonspecific cells (e.g., PBMCs) with high specificity and sensitivity (0.005{\%}). Moreover, TCR analysis data using SERS shows the heterogeneity in the T-cell receptor expression which can inform on the activation status of T-cells and the patient's response to immunotherapy. We believe that this approach may hold potential for numerous applications towards monitoring immune status, understanding therapeutic responses,and effective vaccine development.",
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Dey, S, Vaidyanathan, R, Reza, KK, Wang, J, Wang, Y, Nel, HJ, Law, SC, Tyler, J, Rossjohn, J, Reid, HH, Ibn Sina, AA, Thomas, R & Trau, M 2019, 'A microfluidic-SERSplatform for isolation and immuno-phenotyping of antigen specific T-cells' Sensors and Actuators B: Chemical, vol. 284, pp. 281-288. https://doi.org/10.1016/j.snb.2018.12.099

A microfluidic-SERSplatform for isolation and immuno-phenotyping of antigen specific T-cells. / Dey, Shuvashis; Vaidyanathan, Ramanathan; Reza, K. Kamil; Wang, Jing; Wang, Yuling; Nel, Hendrik J.; Law, Soi Cheng; Tyler, Jennifer; Rossjohn, Jamie; Reid, Hugh H.; Ibn Sina, Abu Ali; Thomas, Ranjeny; Trau, Matt.

In: Sensors and Actuators B: Chemical, Vol. 284, 01.04.2019, p. 281-288.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - A microfluidic-SERSplatform for isolation and immuno-phenotyping of antigen specific T-cells

AU - Dey, Shuvashis

AU - Vaidyanathan, Ramanathan

AU - Reza, K. Kamil

AU - Wang, Jing

AU - Wang, Yuling

AU - Nel, Hendrik J.

AU - Law, Soi Cheng

AU - Tyler, Jennifer

AU - Rossjohn, Jamie

AU - Reid, Hugh H.

AU - Ibn Sina, Abu Ali

AU - Thomas, Ranjeny

AU - Trau, Matt

PY - 2019/4/1

Y1 - 2019/4/1

N2 - T-cells play a major role in host defense mechanisms against many diseases. With the current growth of immunotherapy approaches, there is a strong need for advanced technologies to detect and characterize these immune cells. Herein, we present a simple approach for the isolation of antigen specific T-cells from the complex biological sample based on T-cell receptor (TCR) and peptide major histocompatibility complex (pMHC) interaction. Subsequently, we characterize those antigen specific T-cells by profiling TCR expression heterogeneity. Our approach utilizes an alternating current electrohydrodynamic (ac-EHD) based microfluidic platform for isolation and surface enhanced Raman scattering (SERS) for TCR expression profiling. The use of ac-EHD enables specific isolation of T-cells by generating a nanoscopic shear force at the double layer of the sensing surface which enhances the frequency of pMHC and TCR interactions and consequently shears off the nonspecific targets. TCR expression profiling of the isolated T-cells was performed by encoding them with SERS-labelled pMHCs followed by SERS detection in bulk as well as in single T-Cell. In proof-of-concept experiments, 56.93 ± 7.31% of the total CD4+T-cells were captured from an excess amount of nonspecific cells (e.g., PBMCs) with high specificity and sensitivity (0.005%). Moreover, TCR analysis data using SERS shows the heterogeneity in the T-cell receptor expression which can inform on the activation status of T-cells and the patient's response to immunotherapy. We believe that this approach may hold potential for numerous applications towards monitoring immune status, understanding therapeutic responses,and effective vaccine development.

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KW - Microfluidics

KW - pMHC

KW - SERS

KW - T-cell

KW - TCR

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