Generation and characterization of a human nanobody against VEGFR-2
文献类型：期刊论文
| 作者 | Ma, Lin1; Gu, Kai1,2; Zhang, Cheng-hai1; Chen, Xue-tao1; Jiang, Yi1 ; Melcher, Karsten3; Zhang, Juan2; Wang, Min2; Xu, H. Eric1,3
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| 刊名 | ACTA PHARMACOLOGICA SINICA
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| 出版日期 | 2016-06 |
| 卷号 | 37期号:6页码:857-864 |
| 关键词 | nanobody VEGFR2 anticancer agent anti-angiogenesis phage display |
| ISSN号 | 1671-4083 |
| DOI | 10.1038/aps.2016.2 |
| 文献子类 | Article |
| 英文摘要 | Aim: Nanobody is an antibody fragment consisting of a single monomeric variable antibody domain, which can be used for a variety of biotechnological and therapeutic purposes. The aim of this work was to isolate and characterize a human signal domain antibody against VEGFR-2 domain3 (VEGFR D3) from a phage display library. Methods: To produce antigen-specific recombinant nanobodies with high affinity to VEGFR2 D3, a liquid phase panning strategy was used for all rounds of panning. For nanobody expression and purification, four VEGFR2 D3-blocking clones were subcloned into a pETduet-biotin-MBP expression vector. The recombinant proteins carried an MBP tag to facilitate purification by affinity chromatography. Recombinant NTV(1-4) was obtained after an additional gel filtration chromatography step. The interactions between VEGFR2 D3 and NTV(1-4) were assessed with luminescence-based AlphaScreen assay and SPR assay. Anti-angiogenesis effects were examined in human umbilical vein endothelial cells (HUVECs). Results: In the AlphaScreen assay, NTV1 (100 and 200 nmol/L) elicited the highest binding signal with VEGFR2 D3; NTV2 showed moderate interactions with VEGFR2 D3; NTV3 and NTV4 exhibited little or no interaction with VEGFR2 D3. In the SPR assay, NTV1 displayed a high affinity for VEGFR2 D3 with an equilibrium dissociation constant (K-D) of 49 +/- 1.8 nmol/L. NTV1 (1-1000 nmol/L) dosedependently inhibited the proliferation of HUVECs and the endothelial tube formation by the HUVECs. Conclusion: The nanobody NTV1 is a potential therapeutic candidate for blocking VEGFR2. |
| WOS关键词 | ENDOTHELIAL GROWTH-FACTOR ; BREAST-CANCER ; ANTITUMOR-ACTIVITY ; PHAGE DISPLAY ; ANTIBODY ; RECEPTOR ; THERAPY ; ANGIOGENESIS ; CELLS ; PROLIFERATION |
| 资助项目 | National Natural Science Foundation of China[81072561] ; National Natural Science Foundation of China[81102364] ; National Natural Science Foundation of China[31300607] ; National Natural Science Foundation of China[31230022] |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| CSCD记录号 | CSCD:5722060 |
| WOS记录号 | WOS:000377281000014 |
| 出版者 | ACTA PHARMACOLOGICA SINICA |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/276023]  |
| 专题 | 中国科学院上海药物研究所 |
| 通讯作者 | Ma, Lin; Zhang, Juan; Wang, Min |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Mat Med, VARI SIMM Ctr, Ctr Struct & Funct Drug Targets,Key Lab Receptor, Shanghai 201203, Peoples R China; 2.China Pharmaceut Univ, Sch Life Sci & Technol, State Key Lab Nat Med, Nanjing 210009, Jiangsu, Peoples R China; 3.Van Andel Res Inst, Ctr Struct Biol & Drug Discovery, Lab Struct Sci, 333 Bostwick Ave,NE, Grand Rapids, MI 49503 USA |
| 推荐引用方式 GB/T 7714 | Ma, Lin,Gu, Kai,Zhang, Cheng-hai,et al. Generation and characterization of a human nanobody against VEGFR-2[J]. ACTA PHARMACOLOGICA SINICA,2016,37(6):857-864. |
| APA | Ma, Lin.,Gu, Kai.,Zhang, Cheng-hai.,Chen, Xue-tao.,Jiang, Yi.,...&Xu, H. Eric.(2016).Generation and characterization of a human nanobody against VEGFR-2.ACTA PHARMACOLOGICA SINICA,37(6),857-864. |
| MLA | Ma, Lin,et al."Generation and characterization of a human nanobody against VEGFR-2".ACTA PHARMACOLOGICA SINICA 37.6(2016):857-864. |
入库方式： OAI收割
来源：上海药物研究所
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