ArticlePloS one2018
Soluble T-cell receptor design influences functional yield in an E. coli chaperone-assisted expression system.
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 15 citations in OpenAlex.
- Multi-epitopes vaccine design for surface glycoprotein against SARS-CoV-2 using immunoinformatic approach.Heliyon · 2024Article
- Expression, Purification, and Crystallization of the Vγ9Vδ2 T-cell Receptor Recognizing Protein/Peptide Antigens.The protein journal · 2023Article
- Analysis of CDR3 Sequences from T-Cell Receptor β in Acute Respiratory Distress Syndrome.Biomolecules · 2023Article
- Current and future concepts for the generation and application of genetically engineered CAR-T and TCR-T cells.Frontiers in immunology · 2023Review
- Generalizable design parameters for soluble T cell receptor-based T cell engagers.Journal for immunotherapy of cancer · 2022Article
- Emerging new therapeutic antibody derivatives for cancer treatment.Signal transduction and targeted therapy · 2022Review
- Review
- Stabilization of soluble high-affinity T-cell receptor with de novo disulfide bonds.FEBS letters · 2020Article
- Targeting the MHC Ligandome by Use of TCR-Like Antibodies.Antibodies (Basel, Switzerland) · 2019Review
Corrections and comments
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
There is a quest for production of soluble protein of high quality for the study of T-cell receptors (TCRs), but expression often results in low yields of functional molecules. In this study, we used an E. coli chaperone-assisted periplasmic production system and compared expression of 4 different soluble TCR formats: single-chain TCR (scTCR), two different disulfide-linked TCR (dsTCR) formats, and chimeric Fab (cFab). A stabilized version of scTCR was also included. Additionally, we evaluated the influence of host (XL1-Blue or RosettaBlueTM) and the effect of IPTG induction on expression profiles. A celiac disease patient-derived TCR with specificity for gluten was used, and we achieved detectable expression for all formats and variants. We found that expression in RosettaBlueTM without IPTG induction resulted in the highest periplasmic yields. Moreover, after large-scale expression and protein purification, only the scTCR format was obtained in high yields. Importantly, stability engineering of the scTCR was a prerequisite for obtaining reliable biophysical characterization of the TCR-pMHC interaction. The scTCR format is readily compatible with high-throughput screening approaches that may enable both development of reagents allowing for defined peptide MHC (pMHC) characterization and discovery of potential novel therapeutic leads.
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