Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma
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 v...
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| Format: | Article |
| Language: | English |
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Tsinghua University Press
2009-03-01
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| Series: | Nano Biomedicine and Engineering |
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| Online Access: | https://www.sciopen.com/article/10.5101/nbe.v1i1.p50-56 |
| _version_ | 1797673789439344640 |
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| author | Chenchen Bao Hao Yang Ping Sheng Juxiang Chen Hua Song Xuehua Ding Bin Liu Yicheng Lu Guohan Hu Daxiang Cui |
| author_facet | Chenchen Bao Hao Yang Ping Sheng Juxiang Chen Hua Song Xuehua Ding Bin Liu Yicheng Lu Guohan Hu Daxiang Cui |
| author_sort | Chenchen Bao |
| collection | DOAJ |
| description | 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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| id | doaj.art-6742dc67d94d4ad58b19725c7aebf2e3 |
| institution | Directory Open Access Journal |
| issn | 2150-5578 |
| language | English |
| last_indexed | 2024-03-11T21:50:29Z |
| publishDate | 2009-03-01 |
| publisher | Tsinghua University Press |
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| series | Nano Biomedicine and Engineering |
| spelling | doaj.art-6742dc67d94d4ad58b19725c7aebf2e32023-09-26T09:03:14ZengTsinghua University PressNano Biomedicine and Engineering2150-55782009-03-0111505610.5101/nbe.v1i1.p50-56Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse GliomaChenchen Bao0Hao Yang1Ping Sheng2Juxiang Chen3Hua Song4Xuehua Ding5Bin Liu6Yicheng Lu7Guohan Hu8Daxiang Cui9Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment 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, ChinaDepartment of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment 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, ChinaDepartment of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment 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, ChinaDepartment of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment 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, ChinaHuman 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.https://www.sciopen.com/article/10.5101/nbe.v1i1.p50-56nbeal1gliomacloneexpressionmonoclonal antibody |
| spellingShingle | Chenchen Bao Hao Yang Ping Sheng Juxiang Chen Hua Song Xuehua Ding Bin Liu Yicheng Lu Guohan Hu Daxiang Cui Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma Nano Biomedicine and Engineering nbeal1 glioma clone expression monoclonal antibody |
| title | Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma |
| title_full | Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma |
| title_fullStr | Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma |
| title_full_unstemmed | Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma |
| title_short | Cloning, Expression, Monoclonal Antibody Preparation of Human Gene NBEAL1 and Its Application in Targeted Imaging of Mouse Glioma |
| title_sort | cloning expression monoclonal antibody preparation of human gene nbeal1 and its application in targeted imaging of mouse glioma |
| topic | nbeal1 glioma clone expression monoclonal antibody |
| url | https://www.sciopen.com/article/10.5101/nbe.v1i1.p50-56 |
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