Evolution-guided design of mini-protein for high-contrast in vivo imaging
Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with <sup>99m</sup>Tc, <sup>68</sup>Ga, and <sup>18</sup>F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.
Authors
- Yuquan Wei (ORCID: https://orcid.org/0000-0002-1309-5139)
- Jiadong Yu (ORCID: https://orcid.org/0000-0003-4535-2002)
- Xiaoyan Wang
- Chengcheng Yue (ORCID: https://orcid.org/0000-0001-6093-5615)
- Xiaoqiong Wei (ORCID: https://orcid.org/0000-0001-7576-4820)
- Fulei Zhao (ORCID: https://orcid.org/0000-0002-8866-4041)
- Pei Zhou (ORCID: https://orcid.org/0000-0003-2006-9163)
- Fanlian Zeng
- Wenling Wu (ORCID: https://orcid.org/0000-0001-6244-3489)
- Yawen Hu
- Juan Cheng (ORCID: https://orcid.org/0000-0003-2705-8766)
- Nongyu Huang
- Jiong Li (ORCID: https://orcid.org/0000-0002-3742-630X)
- Xinai Cui
- Chengxin Zhang (ORCID: https://orcid.org/0000-0001-7290-1324)
- Suwen Chen (ORCID: https://orcid.org/0000-0003-2922-9485)
- Guolin Li (ORCID: https://orcid.org/0000-0002-1403-7352)
- Yifan Zhou (ORCID: https://orcid.org/0000-0003-2882-9497)
- Yang Cao (ORCID: https://orcid.org/0000-0001-7052-9346)
- Guangzhou Xiong (ORCID: https://orcid.org/0000-0001-5792-3664)
- Jianyi Yang (ORCID: https://orcid.org/0000-0003-2912-7737)
- Kaijun Cui
Institutions
- National University of Singapore (SG)
- Sichuan University (CN)
- University of Michigan (US)
- West China Hospital of Sichuan University (CN)
- National University Cancer Institute, Singapore (SG)
- State Key Laboratory of Biotherapy
- University of Virginia (US)
- Chengdu University of Traditional Chinese Medicine (CN)
Publication Details
- Journal
- Acta Pharmaceutica Sinica B
- Published
- 2025-07-16
- DOI
- https://doi.org/10.1016/j.apsb.2025.07.015
- Primary Topic
- Cell Image Analysis Techniques
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National University of Singapore
- National Natural Science Foundation of China
- National Science and Technology Major Project
- Natural Science Foundation of Sichuan Province
- National Major Science and Technology Projects of China
- Key Research and Development Program of Sichuan Province