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2 results on '"nucleophosmin mutants"'

1. Monoclonal Antibodies against Nucleophosmin Mutants: Potentials for the Detection of Acute Myeloid Leukemia

Author

Ling Zhang, Shi Tan, Feng-Xian Qin, Hui Chen, Hui-Yuan Shao, Xian-Chun Chen, Hui-Juan Zhang, Xiao-Ming Zhong, Zailin Yang, Li Wang, Yu-Jie Gao, and Liuyang Zhao

Subjects
NPM1, nucleophosmin mutants, medicine.drug_class, Gene mutation, Biology, Monoclonal antibody, law.invention, Mice, law, hemic and lymphatic diseases, medicine, Animals, Humans, acute leukemia, Nucleophosmin, Acute leukemia, Mice, Inbred BALB C, integumentary system, Wild type, Myeloid leukemia, Antibodies, Monoclonal, Nuclear Proteins, General Medicine, Virology, Molecular biology, Immunohistochemistry, Recombinant Proteins, Leukemia, Myeloid, Acute, monoclonal antibody, Mutation, Recombinant DNA, Female, recombinant protein, Research Paper
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.

Published
2011

2. Monoclonal antibodies against nucleophosmin mutants: potentials for the detection of acute myeloid leukemia.

Author

Tan S, Zhang L, Zhong XM, Yang ZL, Zhao LY, Gao YJ, Shao HY, Qin FX, Chen XC, Zhang HJ, Chen H, and Wang L

Subjects
Animals, Antibodies, Monoclonal genetics, Female, Humans, Leukemia, Myeloid, Acute genetics, Mice, Mice, Inbred BALB C, Mutation, Nuclear Proteins genetics, Nuclear Proteins immunology, Nucleophosmin, Recombinant Proteins genetics, Recombinant Proteins immunology, Antibodies, Monoclonal immunology, Immunohistochemistry methods, Leukemia, Myeloid, Acute diagnosis, Nuclear Proteins analysis
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.

Published
2011
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