Preparation and characterization of LGR5 LOOP region-specific nanobodies
文献类型：期刊论文
| 作者 | Jia, Li8; Qiao, Huarui7; Ding, Yuting8; Cui, Qianqian7; Wang, Yingjun8; Geng, Jing8; Tang, Junming4,5,6; Xu, Jianfeng8; Dai, Yuanyuan2,3; Geng, Yong1,7
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| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
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| 出版日期 | 2025-05-01 |
| 卷号 | 229页码:7 |
| 关键词 | LGR5 Nanobody Wnt signaling Topflash |
| ISSN号 | 1046-5928 |
| DOI | 10.1016/j.pep.2025.106680 |
| 英文摘要 | Leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), also known as G-protein-coupled receptor 49 (GPR49), is a class A G-protein-coupled receptor (GPCR) that plays a pivotal role in embryonic development and functions as a marker for adult stem cells in various tissues and organs. LGR5 possesses a large extracellular domain (ecto-domain) enriched with leucine-rich repeats (LRR), primarily responsible for binding to ligands such as R-spondins. The C-terminal LRR extracellular LOOP region of LGR5 refers to the loop structure connecting the C-terminus of LGR5 to the first transmembrane helix. As the LOOP region is located extracellularly, it is readily accessible to exogenous molecules such as antibodies, nanobodies, or small-molecule drugs. In this study, we successfully expressed and purified the LGR5 LOOP region protein in a prokaryotic expression system. The purified protein was subsequently used as an antigen to immunize camels, leading to the generation of nanobodies. These nanobodies are composed solely of the variable domain of the heavy-chain antibody (VHH), with a molecular weight of approximately 15 kDa. Using the purified LGR5 LOOP region protein as an antigen, we isolated nanobodies that specifically bind to it. Subsequent assays demonstrated that the selected nanobody, NB 4C4 and NB 3E8, specifically targeted the LGR5 LOOP region, exhibited an inhibitory effect on (3-cateninmediated Wnt signaling to a certain extent. This study provides insights for the development of LGR5-targeted diagnostic reagents and antibody-based therapeutic strategies. |
| WOS关键词 | HORMONE-RECEPTOR ; ANTIBODY ; CELLS |
| 资助项目 | Shanghai Kailuo Biotechnology Co., Ltd. |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:001422985900001 |
| 出版者 | ACADEMIC PRESS INC ELSEVIER SCIENCE |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/316281]  |
| 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.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Chinese Acad Med Sci, Canc Hosp, Natl Canc Ctr, Natl Clin Res Ctr Canc, Langfang Campus, Langfang 065001, Peoples R China 3.Chinese Acad Med Sci & Peking Union Med Coll, Natl Canc Ctr, Dept Pharm, Natl Clin Res Ctr Canc,Canc Hosp, Beijing 100021, Peoples R China 4.Hubei Univ Med, Inst Biomed, Shiyan 442000, Hubei, Peoples R China 5.Hubei Univ Med, Hubei Key Lab Embryon Stem Cell Res, Shiyan 442000, Hubei, Peoples R China 6.Hubei Univ Med, Fac Basic Med Sci, Dept Physiol, Shiyan 442000, Hubei, Peoples R China 7.Chinese Acad Sci, State Key Lab Drug Res, CAS Key Lab Receptor Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 8.Shanghai Ocean Univ, Coll Food Sci & Technol, Shanghai 201306, Peoples R China |
| 推荐引用方式 GB/T 7714 | Jia, Li,Qiao, Huarui,Ding, Yuting,et al. Preparation and characterization of LGR5 LOOP region-specific nanobodies[J]. PROTEIN EXPRESSION AND PURIFICATION,2025,229:7. |
| APA | Jia, Li.,Qiao, Huarui.,Ding, Yuting.,Cui, Qianqian.,Wang, Yingjun.,...&Geng, Yong.(2025).Preparation and characterization of LGR5 LOOP region-specific nanobodies.PROTEIN EXPRESSION AND PURIFICATION,229,7. |
| MLA | Jia, Li,et al."Preparation and characterization of LGR5 LOOP region-specific nanobodies".PROTEIN EXPRESSION AND PURIFICATION 229(2025):7. |
入库方式： OAI收割
来源：上海药物研究所
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