文献类型：期刊文献

英文题名：Development of a synthetic library of humanized nanobodies for targeted IL-6 inhibition

作者：Wang, Lei[1];Dong, Jiayi[1];Wu, Chenlu[1];Yan, Chenyue[1];Bi, Chong[2];Xu, Chengnan[2];Wu, Yiling[1];Zheng, Wenyun[2];Ma, Xingyuan[1]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, Shanghai, Peoples R China

年份：2024

卷号：12

外文期刊名：FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY

收录：;EI(收录号：20243216817008);WOS:【SCI-EXPANDED(收录号:WOS:001283745200001)】；

基金：The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This study was supported by the National Key Research and Development Project of China (2018YFA0902800), and partially funded by Key Research and Development Project for Logistics Support (20SWAQK24).

语种：英文

外文关键词：nanobody; humanization; stability; synthetic library; phage display; interleukin-6; blocking antibody

摘要：Interleukin-6 (IL-6) is a cytokine that can bind to IL-6 receptor and induce pleiotropic effects. It serves as a critical biomarker, involved in inflammation amplification, tumor progression, and many other disease developments. Nanobodies, featuring small structure and high affinity, are a powerful and versatile tool in medical diagnostics and therapeutics. Here, based on a scaffold optimized for humanization and stability, we developed a synthetic phage display library that rapidly generated high-affinity and humanized nanobodies, negating the need for animal immunization. Using enhanced green fluorescent protein (eGFP) as a benchmark, we demonstrated that the library produced humanized nanobodies with high function and great intracellular stability. The library was then subjected to screening against IL-6. We identified a standout nanobody, NbL3, which exhibited high affinity (22.16 nM) and stability and significantly inhibited IL-6-enhanced migration on the human breast cancer cell MCF-7 at a relatively low concentration. NbL3's strong blocking activity provides a promising therapeutic alternative for the IL-6-targeted intervention strategy, underscoring the broader potential of our synthetic library as a versatile platform for the development of humanized nanobodies against multiple antigens.