Development of a highly specific HER2 monoclonal antibody for immunohistochemistry using protein microarray chips.
Development of a highly specific HER2 monoclonal antibody for immunohistochemistry using protein microarray chips.
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使用蛋白质微阵列芯片开发用于免疫组织化学的高度特异性 HER2 单克隆抗体
DOI：
10.1016/j.bbrc.2017.01.086
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Chen Jian
中科院分区：
文献类型：
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作者：
Qi Lili;Zhou Lixin;Lu Mingmin;Yuan Kehu;Li Zhongwu;Wu Guiyin;Huang Xiaozheng;Shen Yi;Zhao Min;Fu Wei;Chu Boyang;Wang Guangli;Ren Fangfang;Ma Donghui;Chen Jian
HER2是EGFR家族的孤儿受体酪氨酸激酶，被认为是关键的肿瘤驱动基因[1]。HER2扩增的乳腺癌和胃癌可通过其中和抗体Herceptin有效治疗。在临床上，免疫组织化学（IHC）被用作诊断HER2扩增的主要筛查方法[2]。然而，最近的证据表明，经常使用的兔HER2抗体4B5与另一个家族成员HER4交叉反应[3]。4B5免疫组化染色显示正常胃粘膜细胞和部分胃癌组织中有较强的非特异性胞浆和核信号。使用覆盖85%的人类蛋白质组的蛋白质裂解物阵列，我们已经证实了4B5结合HER4和除HER2之外的核蛋白ZSCAN18。非特异性结合解释了正常胃上皮4B5的意外胞质和核染色。最后，我们开发了一种新的小鼠HER2单克隆抗体UMAB36，其对4B5具有相似的敏感性，但仅对蛋白芯片上的17，000种蛋白质中的HER2反应。在129例乳腺癌和158例胃癌样本中，与HER2 FISH参考结果相比，UMAB36显示出100%的灵敏度和特异性，在胃粘膜层中没有非特异性染色。因此，UMAB36可作为检测胃癌人群中HER2扩增的替代高特异性IHC试剂。
HER2 is an orphan receptor tyrosine kinase of the EGFR families and is considered to be a key tumor driver gene [1]. Breast cancer and gastric cancer with HER2 amplification can be effectively treated by its neutralizing antibody, Herceptin. In clinic, Immunohistochemistry (IHC) was used as the primary screening method to diagnose HER2 amplification [2]. However, recent evidence suggested that the frequently used rabbit HER2 antibody 4B5 cross reacted with another family member HER4 [3]. IHC staining with 4B5 also indicated that there was strong non-specific cytoplasmic and nuclear signals in normal gastric mucosal cells and some gastric cancer samples. Using a protein lysate array which covers 85% of the human proteome, we have confirmed that the 4B5 bound to HER4 and a nuclear protein ZSCAN18 besides HER2. The non-specific binding accounts for the unexpected cytoplasmic and nuclear staining of 4B5 of normal gastric epithelium. Finally, we have developed a novel mouse HER2 monoclonal antibody UMAB36 with similar sensitivity to 4B5 but only reacted to HER2 across the 17,000 proteins on the protein chip. In 129 breast cancer and 158 gastric cancer samples, UMAB36 showed 100% sensitivity and specificity comparing to the HER2 FISH reference results with no unspecific staining in the gastric mucosa layer. Therefore, UMAB36 could provide as an alternative highly specific IHC reagent for testing HER2 amplification in gastric cancer populations.