MUC16 is a large transmembrane mucin expressed on the apical surfaces of the epithelium covering the ocular surface, respiratory system and female reproductive tract. The transmembrane mucin is overexpressed by ovarian carcinomas, it is one of the most frequently used diagnostic markers for the disease and it is considered a promising target for immunotherapeutic intervention. Immunodetection of the mucin has to date been through antibodies that recognize its exceptionally large ectodomain. Similar to other membrane anchored mucins, MUC16 has a short cytoplasmic tail (CT), but studies of the biological relevance of the C-terminal domain of MUC16 has been limited by lack of availability of monoclonal antibodies that recognize the native CT. Here, we report the development of a novel monoclonal antibody to the CT region of the molecule that recognizes native MUC16 and its enzymatically released CT region. The antibody is useful for immunoprecipitation of the released CT domain as demonstrated with the OVCAR3 ovarian cancer cell line and can be used for detailed cytolocalization in cells as well as in frozen sections of ocular surface and uterine epithelium.

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针对MUC16细胞质尾巴的单克隆抗体的产生和表征

MUC16是一种大型的跨膜粘蛋白，表达于上皮的顶表面，覆盖眼表，呼吸系统和女性生殖道。跨膜粘蛋白在卵巢癌中过表达，它是该病最常用的诊断标记之一，被认为是免疫治疗干预的有希望的靶标。迄今为止，粘蛋白的免疫检测是通过识别其异常大的胞外域的抗体进行的。与其他膜锚定粘蛋白相似，MUC16具有短的细胞质尾巴（CT），但是由于缺乏可识别天然CT的单克隆抗体，因此对MUC16 C端结构域生物学相关性的研究受到了限制。这里，我们报告了一种新型的单克隆抗体针对分子CT区的发展，该分子识别天然MUC16及其酶促释放的CT区。如OVCAR3卵巢癌细胞系所示，该抗体可用于沉淀的CT域的免疫沉淀，并可用于细胞以及眼表和子宫上皮冰冻切片中的详细细胞定位。