Screening, expression, and functional validation of camelid-derived nanobodies targeting RSPO2
文献类型：期刊论文
| 作者 | Guo, Shaojue2; Zhao, Shuaiying2; Kong, Yingying3,5; Tang, Junming1,6,7; Xu, Jianfeng2; Dai, Yuanyuan8,9; Geng, Yong3,4
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| 刊名 | VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
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| 出版日期 | 2025-05-01 |
| 卷号 | 283页码:8 |
| 关键词 | RSPO2 Wnt signalling Nanobodies Phage display Targeted therapy Specificity |
| ISSN号 | 0165-2427 |
| DOI | 10.1016/j.vetimm.2025.110922 |
| 英文摘要 | Objective: RSPO2 (R-spondin 2) is a key regulator of the Wnt/(3-catenin signaling pathway, involved in embryogenesis, tissue homeostasis, and cancer progression. Despite its therapeutic potential, effective agents targeting RSPO2 remain elusive. To address the unmet need for RSPO2-targeted therapies, we aimed to develop highaffinity nanobodies via phage display and prokaryotic expression, characterizing their binding specificity and functional blockade of RSPO2-LGR4 interactions. This study provides foundational insights into nanobodymediated inhibition of Wnt signaling, supporting future therapeutic strategies against RSPO2-driven pathologies. Methods: Recombinant RSPO2 proteins were constructed and purified using PCR-based recombination. Camels (Camelus bactrianus) were immunized with RSPO2, and phage display was employed to screen nanobody libraries. High-affinity nanobodies were cloned, expressed, purified, and assessed for specificity and binding affinity using biolayer interferometry and protein blotting. Functional validation was performed using TOPFLASH assays to evaluate their impact on Wnt/(3-catenin signaling. Results: Nanobodies with high specificity and nanomolar-range affinity constants (KDs) for RSPO2 were identified. The nanobody effectively inhibited RSPO2-induced Wnt/(3-catenin signaling in human renal epithelial cells. Conclusion: The development of RSPO2-targeting nanobodies offers new prospects for treating RSPO2-related diseases. The nanobody serve as valuable tools for functional research and hold potential as diagnostic and therapeutic agents for RSPO2-driven conditions. |
| WOS关键词 | STRUCTURAL BASIS ; WNT ; RECOGNITION ; ANTIBODY ; PATHWAY |
| WOS研究方向 | Immunology ; Veterinary Sciences |
| 语种 | 英语 |
| WOS记录号 | WOS:001463153700001 |
| 出版者 | ELSEVIER |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/316968]  |
| 专题 | 新药研究国家重点实验室 |
| 通讯作者 | Tang, Junming; Xu, Jianfeng; Dai, Yuanyuan; Geng, Yong |
| 作者单位 | 1.Hubei Univ Med, Fac Basic Med Sci, Dept Physiol, Shiyan 442000, Hubei, Peoples R China 2.Shanghai Ocean Univ, Coll Food Sci & Technol, Dept Biopharmaceut, Shanghai 201306, Peoples R China 3.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China 4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 5.Henan Univ, Coll Pharm, Kaifeng 475000, Henan, Peoples R China 6.Hubei Univ Med, Hubei Key Lab Embryon Stem Cell Res, Shiyan 442000, Hubei, Peoples R China 7.Hubei Univ Med, Inst Biomed, Shiyan 442000, Hubei, Peoples R China 8.Chinese Acad Med Sci & Peking Union Med Coll, Canc Hosp, Natl Canc Ctr, Natl Clin Res Ctr Canc,Dept Pharm, Beijing 100021, Peoples R China 9.Chinese Acad Med Sci, Canc Hosp, Natl Canc Ctr, Natl Clin Res Ctr Canc, Langfang 065001, Peoples R China |
| 推荐引用方式 GB/T 7714 | Guo, Shaojue,Zhao, Shuaiying,Kong, Yingying,et al. Screening, expression, and functional validation of camelid-derived nanobodies targeting RSPO2[J]. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,2025,283:8. |
| APA | Guo, Shaojue.,Zhao, Shuaiying.,Kong, Yingying.,Tang, Junming.,Xu, Jianfeng.,...&Geng, Yong.(2025).Screening, expression, and functional validation of camelid-derived nanobodies targeting RSPO2.VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY,283,8. |
| MLA | Guo, Shaojue,et al."Screening, expression, and functional validation of camelid-derived nanobodies targeting RSPO2".VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY 283(2025):8. |
入库方式： OAI收割
来源：上海药物研究所
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