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Generation of TCR-engineered reference cell samples to control T-cell assay performance
Nicole Bidmon1, Cécile Gouttefangeas2, Sjoerd H van der Burg3
1Translational Oncology at the University Medical Center of the Johannes-Gutenberg University Mainz (TRON gGmbH), Mainz, Germany.
Methods in Enzymology
|January 18, 2020
Summary
A new technology generates T-cell receptor-engineered reference samples (TERS) for standardized in vitro T-cell assays. This innovation improves assay performance control and enables consistent, comparable results across different labs and time points.
Area of Science:
- Immunology
- Cellular Assays
- Biotechnology
Background:
- In vitro cellular assays for antigen-specific T cells are complex and prone to variability.
- Lack of standardized cellular reagents hinders result comparison across experiments and institutions.
Purpose of the Study:
- To develop a robust technology for generating T-cell receptor-engineered reference samples (TERS).
- To enable standardized performance control for key T-cell assays, including MHC-peptide multimer staining, IFN-γ ELISpot, and cytokine flow cytometry.
Main Methods:
- Development of a manufacturing protocol for TERS using electroporation of in vitro-transcribed mRNA encoding stable T-cell receptors.
- Optimization of electroporation settings for versatile in-house production.
Main Results:
- TERS provide defined numbers of antigen-specific T cells for consistent assay performance.
- TERS deliver stable results and allow for long-term storage.
- The optimized protocol facilitates versatile in-house TERS production.
Conclusions:
- TERS technology offers a solution for standardizing complex T-cell assays.
- This approach enhances the reliability and comparability of immunological research data.
- The provided protocol enables accessible, in-house generation of essential reference materials.
