Ayat Hoda, Burrone Oscar R, Sadghizadeh Majid, Jahanzad Eissa, Rastgou Nasrin, Moghadasi Sarrira, Arbabi Mehdi
Department of Genetics, Faculty of Science, Shahrekord University, P.O. Box 115, Shahrekord, Islamic Republic of Iran; Department of Genetics, Faculty of Science, Tarbiat Modaress University, P.O. Box 14115-175, Tehran, Islamic Republic of Iran; National Institute for Genetic Engineering and Biotechnology, P.O. Box 14155-6343, Tehran, Islamic Republic of Iran.
Biologicals. 2013 Nov;41(6):345-54. doi: 10.1016/j.biologicals.2013.05.004. Epub 2013 Jul 8.
Tumor cells expressing HER-2/neu and CEA antigens are potentially ideal targets for antibody-targeted therapy. In this study, two large human combinatorial libraries have been generated from the lymph nodes of breast cancer patients that express HER2 and CEA antigens in their tumors. These 'immune' libraries have been constructed in two different formats of scFv, differing in the length of the peptide linker connecting the two variable VH and VL domains. Libraries derived from these patients may contain a larger pool of anti-tumor antigen antibodies and are useful repertoire for isolating scFvs against any tumor markers. The results of this study showed that we were successful in obtaining human scFvs against HER-2/neu and CEA. For HER-2, cell-panning strategy was performed and resulted in two scFv binders that detected the complete HER-2 receptor on the cell membrane and internalized to the cells. Also, preliminary ELISA data showed that several anti-CEA scFv binders were isolated by panning.
表达HER-2/neu和CEA抗原的肿瘤细胞是抗体靶向治疗的潜在理想靶点。在本研究中，从乳腺癌患者的淋巴结中构建了两个大型人源组合文库，这些患者的肿瘤表达HER2和CEA抗原。这些“免疫”文库以两种不同形式的单链抗体片段（scFv）构建，连接两个可变重链（VH）和轻链（VL）结构域的肽接头长度不同。源自这些患者的文库可能包含更大的抗肿瘤抗原抗体库，是分离针对任何肿瘤标志物的scFv的有用文库。本研究结果表明，我们成功获得了针对HER-2/neu和CEA的人源scFv。对于HER-2，采用细胞淘选策略，得到了两种能在细胞膜上检测完整HER-2受体并内化到细胞内的scFv结合物。此外，初步的ELISA数据表明，通过淘选分离出了几种抗CEA的scFv结合物。
