THETA system allows one-step isolation of tagged proteins through temperature-dependent protein–peptide interaction

• Published in: Communications biology Vol. 2; no. 1; p. 207
• Main Authors: Miura, Kota, Tsuji, Yusuke, Mitsui, Hiromasa, Oshima, Takuya, Noshi, Yosei, Arisawa, Yudai, Okano, Keiko, Okano, Toshiyuki
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 14.06.2019; Nature Publishing Group
• Subjects: 631/1647/664/2229; 631/61/54/1754; 82/1; 82/16; 82/83; Animals; Antibodies, Monoclonal - chemistry; Antibody Affinity; Antibody Specificity; Biocompatible Materials - chemistry; Biology; Biomedical and Life Sciences; Biotinylation; Epitope Mapping; Epitopes - chemistry; External stimuli; Hybridomas - metabolism; Hydrogen-Ion Concentration; Life Sciences; Mice; Mice, Inbred BALB C; Molecular Dynamics Simulation; Monoclonal antibodies; Mutation; Peptides; Peptides - chemistry; Protein interaction; Protein purification; Proteins; Proteins - chemistry; Reproducibility of Results; Temperature; Biomaterials - proteins; Antibody isolation and purification
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-019-0457-8

Tools to control protein-protein interactions by external stimuli have been extensively developed. For this purpose, thermal stimulation can be utilized in addition to light. In this study, we identify a monoclonal antibody termed C13 mAb, which shows an approximately 480-fold decrease in the affinity constant at 37 °C compared to that at 4 °C. Next, we apply this temperature-dependent protein-peptide interaction for one-step protein purifications. We term this THermal-Elution-based TAg system as the THETA system, in which gel-immobilized C13 mAb-derived single-chain variable fragment (scFv) (termed THETAL) is able to bind with proteins tagged by C13 mAb-epitope(s) (THETAS) at 4 °C and thermally release at 37–42 °C. Moreover, to reveal the temperature-dependent interaction mechanism, molecular dynamics simulations are performed along with epitope mapping experiments. Overall, the high specificity and reversibility of the temperature-dependent features of the THETA system will support a wide variety of future applications such as thermogenetics. Kota Miura et al. develop the THermal-Elution-based TAg (THETA) system that allows one-step isolation of tagged proteins. The temperature-dependent protein-peptide interaction used in the THETA system ensures high specificity for protein purification.