Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia
Nucleophosmin (NPM1) gene mutations resulting in cytoplasmic delocalization of Nucleophosmin (NPMc+) are the most common genetic alteration in acute myeloid leukemia (AML). Here, we attempted to prepare monoclonal antibodies (mAbs) against NPM1 mutation A (NPM-mA) and investigated the mAbs' cli...
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| Format: | Online |
| Language: | English |
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Ivyspring International Publisher
2011
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3100738/ |
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pubmed-31007382011-05-24 Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia Tan, Shi Zhang, Ling Zhong, Xiao-Ming Yang, Zai-Lin Zhao, Liu-Yang Gao, Yu-Jie Shao, Hui-Yuan Qin, Feng-Xian Chen, Xian-Chun Zhang, Hui-Juan Chen, Hui Wang, Li Research Paper Nucleophosmin (NPM1) gene mutations resulting in cytoplasmic delocalization of Nucleophosmin (NPMc+) are the most common genetic alteration in acute myeloid leukemia (AML). Here, we attempted to prepare monoclonal antibodies (mAbs) against NPM1 mutation A (NPM-mA) and investigated the mAbs' clinical utility in immunohistochemical detection of NPMc+AML. The pET-32a-NPM-mA vector with the whole open reading frame of the NPM-mA gene was constructed. E.coli BL21 transformed with the vector were induced to express the NPM-mA recombinant protein. BALB/c mice were immunized with the recombinant NPM-mA. Positive clones were selected by indirect ELISA and the mAbs were obtained. Immunohistochemistry was performed to detect the NPMc+ in bone marrow smears from 10 AML patients with NPM-mA. The results showed that the pET-32a-NPM-mA vector was successfully constructed and the NPM-mA recombinant protein was used to immunize the mice. Two positive clones (2G3 and 3F9) were selected. The mAbs against NPM-mA were raised, but did cross-react with wild type NPM1. The mAbs can be used to detect the cytoplasmic dislocation of NPM1 in all AMLs carrying NPM-mA. Our results show that anti-NPM-mA mAbs were produced. Though they would cross-react with wild type NPM1, the mAbs may still have potential in the detection of NPMc+AMLs. Ivyspring International Publisher 2011-05-17 /pmc/articles/PMC3100738/ /pubmed/21611112 Text en © Ivyspring International Publisher. This is an open-access article distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by-nc-nd/3.0/). Reproduction is permitted for personal, noncommercial use, provided that the article is in whole, unmodified, and properly cited. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Tan, Shi Zhang, Ling Zhong, Xiao-Ming Yang, Zai-Lin Zhao, Liu-Yang Gao, Yu-Jie Shao, Hui-Yuan Qin, Feng-Xian Chen, Xian-Chun Zhang, Hui-Juan Chen, Hui Wang, Li |
| spellingShingle |
Tan, Shi Zhang, Ling Zhong, Xiao-Ming Yang, Zai-Lin Zhao, Liu-Yang Gao, Yu-Jie Shao, Hui-Yuan Qin, Feng-Xian Chen, Xian-Chun Zhang, Hui-Juan Chen, Hui Wang, Li Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia |
| author_facet |
Tan, Shi Zhang, Ling Zhong, Xiao-Ming Yang, Zai-Lin Zhao, Liu-Yang Gao, Yu-Jie Shao, Hui-Yuan Qin, Feng-Xian Chen, Xian-Chun Zhang, Hui-Juan Chen, Hui Wang, Li |
| author_sort |
Tan, Shi |
| title |
Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia |
| title_short |
Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia |
| title_full |
Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia |
| title_fullStr |
Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia |
| title_full_unstemmed |
Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia |
| title_sort |
monoclonal antibodies against nucleophosmin mutants: potentials for the detection of acute myeloid leukemia |
| description |
Nucleophosmin (NPM1) gene mutations resulting in cytoplasmic delocalization of Nucleophosmin (NPMc+) are the most common genetic alteration in acute myeloid leukemia (AML). Here, we attempted to prepare monoclonal antibodies (mAbs) against NPM1 mutation A (NPM-mA) and investigated the mAbs' clinical utility in immunohistochemical detection of NPMc+AML. The pET-32a-NPM-mA vector with the whole open reading frame of the NPM-mA gene was constructed. E.coli BL21 transformed with the vector were induced to express the NPM-mA recombinant protein. BALB/c mice were immunized with the recombinant NPM-mA. Positive clones were selected by indirect ELISA and the mAbs were obtained. Immunohistochemistry was performed to detect the NPMc+ in bone marrow smears from 10 AML patients with NPM-mA. The results showed that the pET-32a-NPM-mA vector was successfully constructed and the NPM-mA recombinant protein was used to immunize the mice. Two positive clones (2G3 and 3F9) were selected. The mAbs against NPM-mA were raised, but did cross-react with wild type NPM1. The mAbs can be used to detect the cytoplasmic dislocation of NPM1 in all AMLs carrying NPM-mA. Our results show that anti-NPM-mA mAbs were produced. Though they would cross-react with wild type NPM1, the mAbs may still have potential in the detection of NPMc+AMLs. |
| publisher |
Ivyspring International Publisher |
| publishDate |
2011 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3100738/ |
| _version_ |
1611455435395563520 |