Robust TCR production for the structural study of TCR-pMHC complexes

Oyon-Olea, L.; Dichiara-Rodriguez, M. G.; Hervas-Stubbs, S.; Erausquin, E.; Lopez-Sagaseta, J.

doi:10.1101/2025.05.12.653033

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bioRxiv · 10.1101/2025.05.12.653033

Robust TCR production for the structural study of TCR-pMHC complexes

Oyon-Olea, L.·Dichiara-Rodriguez, M. G.·Hervas-Stubbs, S.·Erausquin, E.·Lopez-Sagaseta, J.

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Abstract

A precise comprehension of how T cell receptors (TCRs) engage their antigens is pivotal for advancing basic research and T cell immunotherapy in cancer. While TCR refolding from inclusion bodies has greatly facilitated X-ray studies over the past decades, the procedures remain labor-intensive and can yield poorly. We have developed a simplified strategy for efficient production of soluble TCRs in CHO cells which, coupled with the removal of N-glycosylation, enable structural studies of TCR-pMHC complexes. An equivalent of just 20 ml of cell culture delivered sufficient deglycosylated TCR (dgTCR) to screen, upon complexation with a cognate pMHC, over 350 crystallization conditions and obtain a high-resolution dataset. This approach illustrates an effective alternative for TCR production to support studies devoted to research and development of TCRs.

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Oyon-Olea, L., Dichiara-Rodriguez, M. G., Hervas-Stubbs, S., Erausquin, E., Lopez-Sagaseta, J.. 2025-05-15. Robust TCR production for the structural study of TCR-pMHC complexes. https://doi.org/10.1101/2025.05.12.653033

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