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Stabilization of soluble high‐affinity T‐cell receptor with de novo disulfide bonds
Author(s) -
Sádio Flávio,
Stadlmayr Gerhard,
Stadlbauer Katharina,
Gräf Maximilian,
Scharrer Agnes,
Rüker Florian,
WozniakKnopp Gordana
Publication year - 2020
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13616
Subject(s) - t cell receptor , mutant , receptor , peptide , chemistry , disulfide bond , t cell , immunoglobulin light chain , biochemistry , microbiology and biotechnology , biology , antibody , immune system , gene , genetics
Soluble T‐cell receptors (TCRs) have recently gained visibility as target‐recognition units of anticancer immunotherapeutic agents. Here, we improved the thermal stability of the well‐expressed high‐affinity A6 TCR by introducing pairs of cysteines in the invariable parts of the α‐ and β‐chain. A mutant with a novel intradomain disulfide bond in each chain also tested superior to the wild‐type in the accelerated stability assay. Binding of the mutant to the soluble cognate peptide (cp)–MHC and to the peptide‐loaded T2 cell line was equal to the wild‐type A6 TCR. The same stabilization motif worked efficiently in TCRs with different specificities, such as DMF5 and 1G4. Altogether, the biophysical properties of the soluble TCR molecule could be improved, without affecting its expression level and antigen‐binding specificity.