单克隆抗体
克洛丹
生物
跨膜蛋白
细胞外
中国仓鼠卵巢细胞
表位
细胞生物学
抗体
计算生物学
细胞培养
遗传学
受体
紧密连接
作者
Hiroshi Ando,Masayo Suzuki,Mariko Kato-Nakano,Shinobu Kawamoto,Hirofumi Misaka,Naoya Kimoto,Akiko Furuya,Kazuyasu Nakamura
标识
DOI:10.1080/09168451.2015.1018124
摘要
Abstract Human claudin-3 (CLDN3) is a tetraspanin transmembrane protein of tight junction structures and is known to be over-expressed in some malignant tumors. Although a specific monoclonal antibody (MAb) against the extracellular domains of CLDN3 would be a valuable tool, generation of such MAbs has been regarded as difficult using traditional hybridoma techniques, because of the conserved sequence homology of CLDN3s among various species. In addition, high sequence similarity is shared among claudin family members, and potential cross-reactivity of MAb should be evaluated carefully. To overcome these difficulties, we generated CLDN3-expressing Chinese hamster ovary and Sf9 cells to use an immunogens and performed cell-based screening to eliminate cross-reactive antibodies. As a result, we generated MAbs that recognized the extracellular loops of CLDN3 but not those of CLDN4, 5, 6, or 9. Further in vitro studies suggested that the isolated MAbs possessed the desired binding properties for the detection or targeting of CLDN3.
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