Development of novel monoclonal antibodies with specific binding affinity for denatured human CD26 in formalin-fixed paraffin-embedded and decalcified specimens
Development of novel monoclonal antibodies with specific binding affinity for denatured human CD26 in formalin-fixed paraffin-embedded and decalcified specimens
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DOI：
10.1371/journal.pone.0218330
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发表时间：
2019-06-13
期刊：
影响因子：
3.7
通讯作者：
Morimoto, Chikao
中科院分区：
文献类型：
--
作者：
Hatano, Ryo;Yamada, Taketo;Morimoto, Chikao
CD26是一种110 kDa的II型跨膜糖蛋白，在其胞外区具有二肽基肽酶IV（DPPIV）活性，具有多种生物学功能，并在炎症反应和肿瘤生物学的调节中发挥重要作用。我们的工作集中在CD26作为一种新的治疗靶点，用于各种肿瘤和免疫疾病，我们最近开发了一种人源化的抗CD26单克隆抗体（mAb），YS 110，它具有良好的安全性和临床活性的恶性胸膜间皮瘤患者。因此，开发一种抗人CD26 mAb，可以在临床环境中清晰可靠地检测福尔马林固定石蜡包埋（FFPE）组织中的变性CD26分子，是至关重要的。为了开发能够结合变性CD26的新型抗CD26 mAb，我们用尿素处理的CD26蛋白免疫小鼠。通过使用一组FFPE人CD26阳性或阴性肿瘤细胞系进行免疫染色，筛选杂交瘤上清液与人CD26的特异性反应性。这种筛选方法使我们能够开发新型抗人CD26 mAb，适用于FFPE非肿瘤和肿瘤组织切片中CD26的免疫组织化学染色，具有可靠的清晰度和强度。具体而言，这些mAb对变性人CD26而不是未变性人CD26显示出强结合亲和力，并且能够检测脱钙标本中的变性人CD26。这些新的抗CD26单克隆抗体可能用于分析骨转移癌症患者的CD26表达，并可能有助于决定未来癌症患者是否适合使用YS 110治疗。
A 110-kDa type II transmembrane glycoprotein with dipeptidyl peptidase IV (DPPIV) activity in its extracellular region, CD26 has a multitude of biological functions and plays an important role in the regulation of inflammatory responses and tumor biology. Our work has focused on CD26 as a novel therapeutic target for various tumors and immune disorders, and we have recently developed a humanized anti-CD26 monoclonal antibody (mAb), YS110, which has promising safety profile and clinical activity in patients with malignant pleural mesothelioma. The development of an anti-human CD26 mAb that can clearly and reliably detect the denatured CD26 molecule in formalin-fixed paraffin-embedded (FFPE) tissues in the clinical setting is therefore of the utmost importance. To develop novel anti-CD26 mAbs capable of binding to denatured CD26, we immunized mice with urea-treated CD26 protein. Hybridoma supernatants were screened for specific reactivity with human CD26 by immunostaining through the use of a set of FFPE human CD26-positive or negative tumor cell lines. This screening method enables us to develop novel anti-human CD26 mAbs suitable for immunohistochemical staining of CD26 in FFPE non-tumor and tumor tissue sections with reliable clarity and intensity. Specifically, these mAbs display strong binding affinity to denatured human CD26 rather than undenatured human CD26, and are capable of detecting denatured human CD26 in decalcified specimens. These novel antiCD26 mAbs are potentially useful for the analysis of CD26 expression in cancer patients with bony metastasis, and may help decide the appropriateness of YS110 therapy for future cancer patients.