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Article | Open Access

Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma

Chenchen Bao1,2, Hao Yang2, Ping Sheng1, Juxiang Chen1, Hua Song2, Xuehua Ding1, Bin Liu2, Yicheng Lu1, Guohan Hu1( ), Daxiang Cui2( )
Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Department of Bio-Nano Science and Engineering, National Key Laboratory of Nano/Micro Fabrication Technology, Key Laboratory for Thin Film and Microfabrication of Ministry of Education, Institute of Micro-Nano Science and Technology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China
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Abstract

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.

Keywords

gliomamonoclonal antibodycloneexpressionNBEAL1

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Nano Biomedicine and Engineering
Volume 1 Issue 1,
March 2009
Pages 50-56
DOI: 10.5101/nbe.v1i1.p50-56
Cite this article:
Bao C, Yang H, Sheng P, et al. Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma. Nano Biomedicine and Engineering, 2009, 1(1): 50-56. https://doi.org/10.5101/nbe.v1i1.p50-56

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Received: 10 November 2009
Accepted: 06 December 2009
Published: 09 December 2009
© 2009 Chenchen Bao et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
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