title: Monoclonal antibodies against nucleophosmin mutants: potentials for the detection of acute myeloid leukemia.
abstract: 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.
authors: 1. Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, 1 Yixueyuan Road, Chongqing, China.
authors: Shi
authors: S
authors: Tan
authors: Ling
authors: L
authors: Zhang
authors: Xiao-Ming
authors: XM
authors: Zhong
authors: Zai-Lin
authors: ZL
authors: Yang
authors: Liu-Yang
authors: LY
authors: Zhao
authors: Yu-Jie
authors: YJ
authors: Gao
authors: Hui-Yuan
authors: HY
authors: Shao
authors: Feng-Xian
authors: FX
authors: Qin
authors: Xian-Chun
authors: XC
authors: Chen
authors: Hui-Juan
authors: HJ
authors: Zhang
authors: Hui
authors: H
authors: Chen
authors: Li
authors: L
authors: Wang
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