A novel monoclonal antibody SMab-2 recognizes endogenous IDH2-R172S of chondrosarcoma
A novel monoclonal antibody SMab-2 recognizes endogenous IDH2-R172S of chondrosarcoma
复制标题
DOI：
10.1016/j.bbrc.2015.02.162
复制
发表时间：
2015-04-17
影响因子：
3.1
通讯作者：
Kato, Yukinari
中科院分区：
文献类型：
--
作者：
Liu, Xing;Ogasawara, Satoshi;Kato, Yukinari
异柠檬酸脱氢酶2 （IDH2）突变在胶质瘤、骨肉瘤、软骨瘤、骨巨细胞瘤和急性髓系白血病中都有报道。虽然IDH2在线粒体中催化异柠檬酸氧化羧化为α -酮戊二酸（α - kg），但突变的IDH2蛋白具有将α - kg转化为肿瘤代谢物R(-)-2-羟基戊二酸（2-HG）的能力。迄今为止，已经建立了几种针对IDH2突变的单克隆抗体（mab），例如针对IDH2- r172k的KMab-1，针对IDH2- r172m的MMab-1和针对IDH2- r172w的WMab-1。虽然多特异性单克隆抗体MsMab-1与IDH2-R172G和IDH2-R172S反应，但尚未建立针对IDH2-R172S的单特异性单克隆抗体。在这项研究中，我们建立了一种新的单克隆抗体SMab-2，它在ELISA中识别IDH2- r172s，但不识别野生型IDH2。虽然SMab-2与大肠杆菌中表达的IDH1-R132S和IDH2-R172S都有反应，但它只与U-2 OS骨肉瘤细胞中表达的IDH2-R172S有反应。此外，在Western blot和免疫细胞化学分析中，SMab-2识别SW1353软骨肉瘤细胞中表达的内源性IDH2-R172S蛋白。SMab-2有望用于诊断携带idh2 - r172s的肿瘤。(C) 2015爱思唯尔公司版权所有。
Isocitrate dehydrogenase 2 (IDH2) mutations have been reported in gliomas, osteosarcomas, cartilaginous tumors, giant cell tumors of bone, and acute myeloid leukemias. Although IDH2 catalyzes the oxidative carboxylation of isocitrate to alpha-ketoglutarate (alpha-KG) in mitochondria, mutated IDH2 proteins possess the ability to change alpha-KG into the oncometabolite R(-)-2-hydroxyglutarate (2-HG). To date, several monoclonal antibodies (mAbs) specific for IDH2 mutations have been established, such as KMab-1 against IDH2-R172K, MMab-1 against IDH2-R172M, and WMab-1 against IDH2-R172W. Although a multi-specific mAb MsMab-1 reacted with IDH2-R172G and IDH2-R172S, a mono-specific mAb against IDH2-R172S has not been established. In this study, we established a novel mAb SMab-2, which recognizes IDH2-R172S but not with wild type IDH2 in ELISA. Although SMab-2 reacted with both IDH1-R132S and IDH2-R172S expressed in Escherichia coli, it reacted with only IDH2-R172S expressed in U-2 OS osteosarcoma cells. Furthermore, SMab-2 recognized endogenous IDH2-R172S protein expressed in SW1353 chondrosarcoma cells in Western blot and immunocytochemical analyses. SMab-2 is expected to be useful for diagnosis of IDH2-R172S-bearing tumors. (C) 2015 Elsevier Inc. All rights reserved.