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

Chenchen Bao; Hao Yang; Ping Sheng; Juxiang Chen; Hua Song; Xuehua Ding; Bin Liu; Yicheng Lu; Guohan Hu; Daxiang Cui

doi:10.5101/nbe.v1i1.p50-56

Nano Biomedicine and Engineering (Mar 2009)

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

Chenchen Bao,
Hao Yang,
Ping Sheng,
Juxiang Chen,
Hua Song,
Xuehua Ding,
Bin Liu,
Yicheng Lu,
Guohan Hu,
Daxiang Cui

Affiliations

Chenchen Bao: Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Hao Yang: 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
Ping Sheng: Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Juxiang Chen: Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Hua Song: 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
Xuehua Ding: Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Bin Liu: 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
Yicheng Lu: Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Guohan Hu: Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
Daxiang Cui: 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

DOI: https://doi.org/10.5101/nbe.v1i1.p50-56
Journal volume & issue: Vol. 1, no. 1
pp. 50 – 56

Abstract

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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.

Published in Nano Biomedicine and Engineering

ISSN: 2097-3837 (Print); 2150-5578 (Online)
Publisher: Tsinghua University Press
Country of publisher: China
LCC subjects: Science: Biology (General); Medicine
Website: https://www.sciopen.com/journal/2150-5578

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