Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma
Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma
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人基因NBEAL1的克隆、表达、单克隆抗体制备及其在小鼠胶质瘤靶向成像中的应用
DOI：
10.5101/nbe.v1i1.p50-56
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发表时间：
2009-12
期刊：
影响因子：
--
通讯作者：
胡国汉
中科院分区：
文献类型：
--
作者：
胡国汉
人神经海滩蛋白样1（NBEAL 1）基因是BEACH-WD 40结构域家族的重要成员，在I期胶质瘤中表达明显。本研究从U251细胞中提取总RNA，通过RT-PCR获得其cDNA序列，并将部分NBEAL 1 cDNA片段克隆到pGEX-KG载体中。重组表达载体在大肠杆菌BL 21中获得了GST融合蛋白的高效表达。通过亲和层析纯化NBEAL 1重组蛋白。制备了抗重组NBEAL 1蛋白的单克隆抗体。Western Blotting分析鉴定其生物活性。抗NBEAL 1抗体与CdTe量子点缀合。将所得的抗NBEAL 1抗体缀合的纳米探针经由尾血管注射到小鼠中。12 h后，IVIS成像系统可清晰观察到制备的纳米探针定位于胶质瘤模型小鼠脑组织中。结论：成功表达了NBEAL 1蛋白，并制备了其单克隆抗体。抗NBEAL 1抗体缀合的量子点可用于对神经胶质瘤成像。这些制备的纳米探针在胶质瘤的早期检测中具有很大的潜力。
Human Neurobeachin-like 1(NBEAL1) gene was an important member of BEACH-WD40 domain family, which was confirmed to be overexpressed in I stage glioma. In this study, we extracted total RNAs from U251 cell line, acquired its cDNA sequence by RT-PCR, and cloned part of NBEAL1 cDNA fragments into the vector pGEX-KG. The recombinant expression vector achieved high expression in E.coli BL21 as a GST fusion protein. NBEAL1 recombinant protein was purified by affinity chromatography. Monoclonal antibody was prepared against the recombinant NBEAL1 protein. Its bioactivity was identified by Western Blotting analysis. Anti-NBEAL1 antibody was conjugated with CdTe quantum dots. Resultant anti-NBEAL1 antibody-conjugated nanoprobes were injected into mice via tail vessel. After 12h, it is clearly observed that prepared nanoprobes located in brain tissues of mice model with glioma by IVIS Imaging system. In conclusion, NBEAL1 protein was successfully expressed and its monoclonal antibody was successfully prepared. Anti-NBEAL1 antibody-conjugated quantum dots may be used to image glioma. These prepared nanoprobes have great potential in early detection of glioma.
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影响因子：
64.8
作者：
Lu, W;Ji, ZQ;Rimberg, AJ
通讯作者：
Rimberg, AJ
影响因子：
5.3
作者：
Su, YH;Balice-Gordon, RJ;Cooke, NE
通讯作者：
Cooke, NE
DOI：
10.1523/jneurosci.20-23-08551.2000
发表时间：
2000-12
期刊：
The Journal of Neuroscience
影响因子：
--
作者：
Xiaolu Wang;F. Herberg;M. Laue;C. Wüllner;Bin Hu;E. Petrasch‐Parwez;M. Kilimann
通讯作者：
Xiaolu Wang;F. Herberg;M. Laue;C. Wüllner;Bin Hu;E. Petrasch‐Parwez;M. Kilimann
影响因子：
17.3
作者：
E G L Bywaters
通讯作者：
E G L Bywaters
影响因子：
4.1
作者：
Galanaud, D;Chinot, O;Cozzone, PJ
通讯作者：
Cozzone, PJ