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Development of an RNA-based kit for easy generation of TCR-engineered lymphocytes to control T-cell assay performance

Development of an RNA-based kit for easy generation of TCR-engineered lymphocytes to control T-cell assay performance
Development of an RNA-based kit for easy generation of TCR-engineered lymphocytes to control T-cell assay performance

Cell-based assays to monitor antigen-specific T-cell responses are characterized by their high complexity and should be conducted under controlled conditions to lower multiple possible sources of assay variation. However, the lack of standard reagents makes it difficult to directly compare results generated in one lab over time and across institutions. Therefore TCR-engineered reference samples (TERS) that contain a defined number of antigen-specific T cells and continuously deliver stable results are urgently needed. We successfully established a simple and robust TERS technology that constitutes a useful tool to overcome this issue for commonly used T-cell immuno-assays. To enable users to generate large-scale TERS, on-site using the most commonly used electroporation (EP) devices, an RNA-based kit approach, providing stable TCR mRNA and an optimized manufacturing protocol were established. In preparation for the release of this immuno-control kit, we established optimal EP conditions on six devices and initiated an extended RNA stability study. Furthermore, we coordinated on-site production of TERS with 4 participants. Finally, a proficiency panel was organized to test the unsupervised production of TERS at different laboratories using the kit approach. The results obtained show the feasibility and robustness of the kit approach for versatile in-house production of cellular control samples.

Immunoguiding, Immunomonitoring, Performance, Standardization, T-cell assays, Validation
0022-1759
74-82
Bidmon, Nicole
63ab481b-2caa-41f9-a900-fd7f37cd1376
Kind, Sonja
b691d96a-a703-4d8f-99f1-de694c28570d
Welters, Marij J.P.
ece995e7-8d4c-431d-83a9-fc0a24f6fdf4
Joseph-Pietras, Deborah
3002f7ea-3689-40c3-8949-9da8f0753c91
Laske, Karoline
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Maurer, Dominik
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Hadrup, Sine Reker
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Schreibelt, Gerty
b9e389e6-0ca5-4172-9065-917f10ad4403
Rae, Richard
c207b53e-18dd-4912-ba7f-447729d1670a
Sahin, Ugur
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Gouttefangeas, Cécile
e06644af-a88f-4d60-b11c-debfaffefd0d
Britten, Cedrik M.
a303deac-ffba-4a55-9a7e-fa5e158c0f8d
van der Burg, Sjoerd H.
1e880617-4966-4046-b70c-0ef451c2c2ab
Bidmon, Nicole
63ab481b-2caa-41f9-a900-fd7f37cd1376
Kind, Sonja
b691d96a-a703-4d8f-99f1-de694c28570d
Welters, Marij J.P.
ece995e7-8d4c-431d-83a9-fc0a24f6fdf4
Joseph-Pietras, Deborah
3002f7ea-3689-40c3-8949-9da8f0753c91
Laske, Karoline
8cdca8cb-bf0c-415f-8eb5-51acd6c65f47
Maurer, Dominik
2d9f2343-cc3e-476b-87e0-9c7004549fd1
Hadrup, Sine Reker
6d51f8ef-0ef2-44a8-8383-19e29d2dc041
Schreibelt, Gerty
b9e389e6-0ca5-4172-9065-917f10ad4403
Rae, Richard
c207b53e-18dd-4912-ba7f-447729d1670a
Sahin, Ugur
153fa486-fe00-40ac-9d70-7e049091a991
Gouttefangeas, Cécile
e06644af-a88f-4d60-b11c-debfaffefd0d
Britten, Cedrik M.
a303deac-ffba-4a55-9a7e-fa5e158c0f8d
van der Burg, Sjoerd H.
1e880617-4966-4046-b70c-0ef451c2c2ab

Bidmon, Nicole, Kind, Sonja, Welters, Marij J.P., Joseph-Pietras, Deborah, Laske, Karoline, Maurer, Dominik, Hadrup, Sine Reker, Schreibelt, Gerty, Rae, Richard, Sahin, Ugur, Gouttefangeas, Cécile, Britten, Cedrik M. and van der Burg, Sjoerd H. (2018) Development of an RNA-based kit for easy generation of TCR-engineered lymphocytes to control T-cell assay performance. Journal of Immunological Methods, 458, 74-82. (doi:10.1016/j.jim.2018.04.007).

Record type: Article

Abstract

Cell-based assays to monitor antigen-specific T-cell responses are characterized by their high complexity and should be conducted under controlled conditions to lower multiple possible sources of assay variation. However, the lack of standard reagents makes it difficult to directly compare results generated in one lab over time and across institutions. Therefore TCR-engineered reference samples (TERS) that contain a defined number of antigen-specific T cells and continuously deliver stable results are urgently needed. We successfully established a simple and robust TERS technology that constitutes a useful tool to overcome this issue for commonly used T-cell immuno-assays. To enable users to generate large-scale TERS, on-site using the most commonly used electroporation (EP) devices, an RNA-based kit approach, providing stable TCR mRNA and an optimized manufacturing protocol were established. In preparation for the release of this immuno-control kit, we established optimal EP conditions on six devices and initiated an extended RNA stability study. Furthermore, we coordinated on-site production of TERS with 4 participants. Finally, a proficiency panel was organized to test the unsupervised production of TERS at different laboratories using the kit approach. The results obtained show the feasibility and robustness of the kit approach for versatile in-house production of cellular control samples.

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Accepted/In Press date: 18 April 2018
e-pub ahead of print date: 21 April 2018
Keywords: Immunoguiding, Immunomonitoring, Performance, Standardization, T-cell assays, Validation

Identifiers

Local EPrints ID: 421132
URI: https://eprints.soton.ac.uk/id/eprint/421132
ISSN: 0022-1759
PURE UUID: 89711df6-0d84-434f-8ca0-34b39af66125

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Date deposited: 23 May 2018 16:30
Last modified: 16 Jul 2019 17:16

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Contributors

Author: Nicole Bidmon
Author: Sonja Kind
Author: Marij J.P. Welters
Author: Deborah Joseph-Pietras
Author: Karoline Laske
Author: Dominik Maurer
Author: Sine Reker Hadrup
Author: Gerty Schreibelt
Author: Richard Rae
Author: Ugur Sahin
Author: Cécile Gouttefangeas
Author: Cedrik M. Britten
Author: Sjoerd H. van der Burg

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