Identification and characterization of tumor antigens by using antibody phage display and intrabody strategies
Identification and characterization of tumor antigens by using antibody phage display and intrabody strategies
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DOI：
10.1016/j.molimm.2007.03.023
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发表时间：
2007-07-01
影响因子：
3.6
通讯作者：
Poul, Marie-Alix
中科院分区：
文献类型：
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作者：
Goenaga, Anne-Laure;Zhou, Yu;Poul, Marie-Alix
为了生成一组结合人类乳腺癌的抗体，选择了人类单链 Fv 噬菌体展示库来快速内化到 SK-BR-3 乳腺癌细胞系中。在所研究的 55 种细胞结合抗体中鉴定出了 13 种独特的抗体，与正常上皮细胞相比，所有这些抗体都显示出肿瘤细胞的特异性染色。其中两种抗体结合 ErbB2 癌基因，而 6 种抗体结合肿瘤标志物转铁蛋白受体 (TfR)。通过开发 scFv 免疫沉淀方法，我们能够使用 LC-MS/MS 来鉴定与其中一种抗体 (3GA5) 结合的抗原，即 FPRP（前列腺素 F2α 受体调节蛋白）/EWI-F/CD9P-1（CD9 伴侣 1），这是一种 Ig 超家族成员，已被描述为直接与 CD9 和 CD81 四跨膜蛋白相互作用，并在贴壁癌细胞系中过度表达。尽管3GA5 scFv没有直接的抗增殖作用，但scFv的细胞内表达能够敲低CD9P-1的表达，并且可用于进一步确定四跨膜蛋白系统在增殖和转移中的作用。此外，3GA5 scFv 能快速内化到乳腺肿瘤细胞中，并具有将细胞毒性药物靶向递送至乳腺癌的潜力。这项研究从原理上证明了直接选择肿瘤细胞上的噬菌体抗体库可以有效地导致相关肿瘤标志物的鉴定和功能表征。 (C) 2007 Elsevier Ltd. 保留所有权利。
To generate a panel of antibodies binding human breast cancers, a human single chain Fv phage display library was selected for rapid internalization into the SK-BR-3 breast cancer cell line. Thirteen unique antibodies were identified within the 55 cell binding antibodies studied, all of them showing specific staining of tumor cells compare to normal epithelial eel Is. Two of the antibodies bound the ErbB2 oncogene while 6 bound the tumor marker transferrin receptor (TfR). By developing a scFv immunoprecipitation method, we were able to use LC-MS/MS to identify the antigen bound by one of the antibodies (3GA5) as FPRP (prostaglandin F2alpha receptor-regulatory protein)/EWI-F/CD9P-1 (CD9 partner 1) an Ig superfamily member that has been described to interact directly with CD9 and CD81 tetraspanins and to be overexpressed in adherent cancer cell lines. Although the 3GA5 scFv had no direct anti-proliferative effect, intracellular expression of the scFv was able to knockdown CD9P-1 expression and could be used to further define the role of the tetraspanin system in proliferation and metastasis. Moreover, the 3GA5 scFv was rapidly internalized into breast tumor cells and could have potential for the targeted delivery of cytotoxic agents to breast cancers. This study is the proof of principle that the direct selection of phage antibody libraries on tumor cells can effectively lead to the identification and functional characterization of relevant tumor markers. (C) 2007 Elsevier Ltd. All rights reserved.