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Screening and preparation of nanobodies for SIGLEC-15 detection

文献类型：期刊论文

作者Zhao, Shuaiying6,7; Li, Lingyun1,6; Lan, Zhongyun6; Hou, Xiangyun5,6; Huang, Ruimin5,6; Jin, Quanfang4; Huang, Qiting6,7; Jia, Li6,7; Kong, Yingying6; Wen, Jianchuan6,7
刊名PROTEIN EXPRESSION AND PURIFICATION
出版日期2025-05-01
卷号229页码:9
关键词SIGLEC-15 Nanobody Expression Purification
ISSN号1046-5928
DOI10.1016/j.pep.2025.106679
英文摘要As an important member of the Siglec family, SIGLEC-15 plays an important role in osteoclast differentiation, bone remodeling, and tumor immune evasion. In the tumor microenvironment, SIGLEC-15 functions independently of the B7-H1/PD-1 pathway. In this study, the SIGLEC-15 fusion protein (SIGLEC-15-Fc) was successfully expressed and purified using a eukaryotic expression system. This protein was then used as an antigen to immunize camelids, inducing the production of specific nanobodies (VHHs) targeting SIGLEC-15. The resulting nanobodies exhibited a molecular weight of approximately 15 kDa. After screening, we identified two nanobody strains, Nb1C8 and Nb2D7, both of which bind SIGLEC-15 without competition. We confirmed the nanobodies' high affinity and stability through Octec platform and stability analyses. Flow cytometry analysis demonstrated that Nb1C8 and Nb2D7 specifically binds to SIGLEC-15 which naturally expressed on bladder cancer cells. This study marks the first development of nanobodies specifically targeting SIGLEC-15, providing a solid foundation for the development of SIGLEC-15-related diagnostic tools and antibody therapeutics.
WOS关键词THERAPY ; GENERATION ; TARGET ; TUMOR
WOS研究方向Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种英语
WOS记录号WOS:001421464700001
出版者ACADEMIC PRESS INC ELSEVIER SCIENCE
源URL[http://119.78.100.183/handle/2S10ELR8/316302]  
专题新药研究国家重点实验室
通讯作者Li, Lingyun; Geng, Yong; Xu, Jianfeng; Dai, Yuanyuan
作者单位1.Chinese Acad Sci, Shenzhen Inst Adv Technol, Ctr Cognit Technol, Shenzhen 518055, Guangdong, Peoples R China
2.Chinese Acad Med Sci, Natl Canc Ctr, Natl Clin Res Ctr Canc, Canc Hosp, Langfang campus, Langfang 065001, Peoples R China
3.Chinese Acad Med Sci & Peking Union Med Coll, Canc Hosp, Natl Canc Ctr, Dept Pharm,Natl Clin Res Ctr Canc, Beijing 100021, Peoples R China
4.Shanghai Baoshan Hosp Integrated Tradit Chinese &, Shanghai, Peoples R China
5.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
6.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China
7.Shanghai Ocean Univ, Coll Food Sci & Technol, Shanghai 201306, Peoples R China
推荐引用方式
GB/T 7714		 Zhao, Shuaiying,Li, Lingyun,Lan, Zhongyun,et al. Screening and preparation of nanobodies for SIGLEC-15 detection[J]. PROTEIN EXPRESSION AND PURIFICATION,2025,229:9.
APA Zhao, Shuaiying.,Li, Lingyun.,Lan, Zhongyun.,Hou, Xiangyun.,Huang, Ruimin.,...&Dai, Yuanyuan.(2025).Screening and preparation of nanobodies for SIGLEC-15 detection.PROTEIN EXPRESSION AND PURIFICATION,229,9.
MLA Zhao, Shuaiying,et al."Screening and preparation of nanobodies for SIGLEC-15 detection".PROTEIN EXPRESSION AND PURIFICATION 229(2025):9.

入库方式： OAI收割

来源：上海药物研究所

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