Establishment of Anti-Human ATRX Monoclonal Antibody AMab-6.
Establishment of Anti-Human ATRX Monoclonal Antibody AMab-6.
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DOI：
10.1089/mab.2016.0037
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发表时间：
2016-10
影响因子：
--
通讯作者：
Kato Y
中科院分区：
文献类型：
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作者：
Ogasawara S;Fujii Y;Kaneko MK;Oki H;Sabit H;Nakada M;Suzuki H;Ichimura K;Komori T;Kato Y
神经胶质瘤是最常见的脑肿瘤，具有异质分子背景。与传统的组织学分类相比，神经胶质瘤的分子亚组更能将患者分为不同的组。弥漫性胶质瘤亚型诊断最重要的分子是异柠檬酸脱氢酶（IDH）、TERT启动子和α-地中海贫血/智力迟钝综合征-X连锁（ATRX）突变和1p/19q的联合缺失。其中，IDH和ATRX突变可以使用特异性单克隆抗体（mAb）进行诊断。我们开发了许多针对IDH突变体的单克隆抗体，包括针对IDH1-R132H突变体的HMab-1/HMab-2和针对IDH1/2突变体的多特异性单克隆抗体MsMab-1/MsMab-2。相比之下，针对 ATRX 的高灵敏度单克隆抗体仍有待建立。在这项研究中，我们用重组人 ATRX 免疫小鼠，并开发了一种新型单克隆抗体 AMab-6。 AMab-6的解离常数测定为9.7 × 10−10 M，表明AMab-6的结合亲和力非常高。此外，AMab-6 在蛋白质印迹和免疫组织化学分析中灵敏地检测 ATRX，表明 AMab-6 可以成为免疫组织化学分析中确定胶质瘤 ATRX 突变状态的标准标记。
Gliomas are the most frequently occurring brain tumors with a heterogeneous molecular background. The molecular subgrouping of gliomas more prognostically stratifies patients into distinct groups compared with conventional histological classification. The most important molecules for the subtype diagnosis of diffuse gliomas are mutations of isocitrate dehydrogenase (IDH), TERT promoter, and α-thalassemia/mental-retardation-syndrome-X-linked (ATRX) and the codeletion of 1p/19q. Among them, IDH and ATRX mutations can be diagnosed using specific monoclonal antibodies (mAbs). We have developed many mAbs against IDH mutants, including HMab-1/HMab-2 against IDH1-R132H and multispecific mAbs MsMab-1/MsMab-2 against IDH1/2 mutations. In contrast, highly sensitive mAbs against ATRX remain to be established. In this study, we immunized mice with recombinant human ATRX and developed a novel mAb, AMab-6. The dissociation constant of AMab-6 was determined to be 9.7 × 10−10 M, indicating that the binding affinity of AMab-6 is very high. Furthermore, AMab-6 sensitively detects ATRX in Western blot and immunohistochemical analyses, indicating that AMab-6 could become the standard marker to determine the ATRX mutation status of gliomas in immunohistochemical analyses.