Monoclonal antibody (mAb) technology is an excellent tool for the discovery of overexpressed cell surface tumour antigens and the development of targeting agents. Here, we report the development of two novel mAbs against CFPAC-1 human pancreatic cancer cells. Using ELISA, flow cytometry, immunoprecipitation, mass spectrometry, Western blot and immunohistochemistry, we found that the target antigens recognised by the two novel mAbs KU44.22B and KU44.13A, are integrin α3 and CD26 respectively, with high levels of expression in human pancreatic and other cancer cell lines and human pancreatic cancer tissue microarrays. Treatment with naked anti-CD26 mAb KU44.13A did not have any effect on the growth and migration of cancer cells nor did it induce receptor downregulation. In contrast, treatment with anti-integrin α3 mAb KU44.22B inhibited growth in vitro of Capan-2 cells, increased migration of BxPC-3 and CFPAC-1 cells and induced antibody internalisation. Both novel mAbs are capable of detecting their target antigens by immunohistochemistry but not by Western blot. These antibodies are excellent tools for studying the role of integrin α3 and CD26 in the complex biology of pancreatic cancer, their prognostic and predictive values and the therapeutic potential of their humanised and/or conjugated versions in patients whose tumours overexpress integrin α3 or CD26.

中文翻译：

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两种新型抗人胰腺癌CD26和整联蛋白α3的新型单克隆抗体的开发和应用。

单克隆抗体（mAb）技术是发现过表达的细胞表面肿瘤抗原和开发靶向剂的出色工具。在这里，我们报道了针对CFPAC-1人胰腺癌细胞的两种新型mAb的开发。使用ELISA，流式细胞仪，免疫沉淀，质谱，Western印迹和免疫组织化学，我们发现被两种新型单克隆抗体KU44.22B和KU44.13A识别的靶抗原分别是整联蛋白α3和CD26，在人类中高水平表达胰腺和其他癌细胞系以及人类胰腺癌组织微阵列。裸露的抗CD26 mAb KU44.13A处理对癌细胞的生长和迁移没有任何影响，也不会诱导受体下调。相反，用抗整联蛋白α3mAb KU44处理。22B抑制Capan-2细胞的体外生长，增加BxPC-3和CFPAC-1细胞的迁移并诱导抗体内在化。两种新型mAb均能够通过免疫组织化学而非Western印迹检测其靶抗原。这些抗体是研究整联蛋白α3和CD26在胰腺癌的复杂生物学中的作用，其预后和预测价值以及其人源化和/或缀合形式对肿瘤过度表达整联蛋白α3或CD26的患者的治疗潜力的出色工具。