Christian Doppler Laboratory for Innovative Immunotherapeutics, Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria.
FEBS Lett. 2020 Feb;594(3):477-490. doi: 10.1002/1873-3468.13616. Epub 2019 Oct 8.
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.
可溶性 T 细胞受体（TCRs）最近作为抗癌免疫治疗药物的靶识别单位受到关注。在这里，我们通过在 α 和 β 链的不变部分引入一对半胱氨酸来提高表达良好的高亲和力 A6 TCR 的热稳定性。一种在每条链中具有新型的域内二硫键的突变体在加速稳定性测定中也表现优于野生型。突变体与可溶性同源肽（cp）-MHC 以及负载肽的 T2 细胞系的结合与野生型 A6 TCR 相当。相同的稳定化模体在具有不同特异性的 TCR 中（如 DMF5 和 1G4）同样有效。总的来说，可溶性 TCR 分子的生物物理特性可以得到改善，而不会影响其表达水平和抗原结合特异性。
