Establishment of a novel monoclonal antibody SMab-1 specific for IDH1-R132S mutation
Establishment of a novel monoclonal antibody SMab-1 specific for IDH1-R132S mutation
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DOI：
10.1016/j.bbrc.2011.02.102
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发表时间：
2011-03-25
影响因子：
3.1
通讯作者：
Kato, Yukinari
中科院分区：
文献类型：
--
作者：
Kaneko, Mika Kato;Tian, Wei;Kato, Yukinari
异柠檬酸脱氢酶1（IDH 1）突变是胶质瘤中早期和频繁的遗传改变，其特异于IDH 1中保守且功能重要的精氨酸132（R132）中的单个密码子。我们早期建立了一种单克隆抗体（mAb），IMab-1，它对含R132 H的IDH 1（IDH 1-R132 H）具有特异性，IDH 1是神经胶质瘤中最常见的IDH 1突变。为了建立IDH 1-R132 S特异性mAb，我们用含有R132 S的IDH 1（IDH 1-R132 S）肽免疫小鼠。细胞融合后用仙台病毒包膜。在ELISA中筛选IDH 1-R132 S特异性mAb。一种mAb SMab-1与IDH 1-R132 S肽反应，但不与其他IDH 1突变体反应。Western-blot分析表明，SMab-1仅与IDH 1-R132 S蛋白反应，而不与IDH 1-WT蛋白或IDH 1突变体反应，表明SMab-1是IDH 1-R132 S特异性的。此外，SMab-1在免疫细胞化学和免疫组织化学中特异性染色表达IDH 1-R132 S的胶质母细胞瘤细胞，但不与含有IDH 1-WT或IDH 1-R132 H的胶质母细胞瘤细胞反应。我们新建立了一种抗IDH 1-R132 S特异性mAb SMab-1，用于诊断携带突变的胶质瘤。(C)2011 Elsevier Inc. All rights reserved.
Isocitrate dehydrogenase 1 (IDH1) mutations, which are early and frequent genetic alterations in gliomas, are specific to a single codon in the conserved and functionally important Arginine 132 (R132) in IDH1. We earlier established a monoclonal antibody (mAb), IMab-1, which is specific for R132H-containing IDH1 (IDH1-R132H), the most frequent IDH1 mutation in gliomas. To establish IDH1-R132S-specific mAb, we immunized mice with R132S-containing IDH1 (IDH1-R132S) peptide. After cell fusion using Sendai virus envelope. IDH1-R132S-specific mAbs were screened in ELISA. One mAb, SMab-1, reacted with the IDH1-R132S peptide, but not with other IDH1 mutants. Western-blot analysis showed that SMab-1 reacted only with the IDH1-R132S protein, not with IDH1-WT protein or IDH1 mutants, indicating that SMab-1 is IDH1-R132S-specific. Furthermore, SMab-1 specifically stained the IDH1-R132S-expressing glioblastoma cells in immunocytochemistry and immunohistochemistry, but did not react with IDH1-WT or IDH1-R132H-containing glioblastoma cells. We newly established an anti-IDH1-R132S-specific mAb SMab-1 for use in diagnosis of mutation-bearing gliomas. (C) 2011 Elsevier Inc. All rights reserved.