Sithambaram Devilakshmi, Palanivelu Shanthi, Subramanian Krishnakumar, Sahoo Sanjeeb, Verma Rama S
Department of Biotechnology, Indian Institute of Technology Madras, Chennai, India.
Hybridoma (Larchmt). 2011 Dec;30(6):511-8. doi: 10.1089/hyb.2011.0069.
Epithelial cell adhesion molecule (Ep-CAM) is a 40 kDa transmembrane glycoprotein overexpressed in majority of tumor epithelial cells and has a major morphoregulatory function, relevant not only to epithelial tissue development, but also in carcinogenesis and tumor progression. Since Ep-CAM localizes at the cell surface of most carcinomas, the molecule is an attractive target for immunotherapy and several strategies have been deployed to treat cancer using Ep-CAM targeting, including MAb therapy. For improving effective targeting of this protein for diagnostics in various clinical samples, we generated and characterized an anti-Ep-CAM MAb (C4) using recombinant Ep-CAM protein, comprising the highly immunogenic domain. The specificity of C4-MAb was characterized in Ep-CAM positive cell lines (PC3 and MCF-7) by flow cytometry and immunofluorescence. The immunohistochemistry analysis in clinical tissue samples showed specific detection of epithelial antigens in breast, colon, stomach, and prostate carcinomas. Thus, this Ep-CAM MAb (C4-MAb) could be used for both diagnostic and therapeutic applications due to its specificity.
上皮细胞粘附分子(Ep-CAM)是一种40 kDa的跨膜糖蛋白,在大多数肿瘤上皮细胞中过表达,具有主要的形态调节功能,不仅与上皮组织发育有关,还与致癌作用和肿瘤进展有关。由于Ep-CAM定位于大多数癌的细胞表面,该分子是免疫治疗的一个有吸引力的靶点,并且已经采用了几种策略来使用Ep-CAM靶向治疗癌症,包括单克隆抗体治疗。为了改善该蛋白在各种临床样本中用于诊断的有效靶向性,我们使用包含高免疫原性结构域的重组Ep-CAM蛋白制备并鉴定了一种抗Ep-CAM单克隆抗体(C4)。通过流式细胞术和免疫荧光在Ep-CAM阳性细胞系(PC3和MCF-7)中对C4单克隆抗体的特异性进行了鉴定。临床组织样本的免疫组织化学分析显示在乳腺癌、结肠癌、胃癌和前列腺癌中可特异性检测上皮抗原。因此,这种Ep-CAM单克隆抗体(C4单克隆抗体)因其特异性可用于诊断和治疗应用。