Morita Shinkichi, Mochizuki Mai, Shibuya-Takahashi Rie, Nakamura-Shima Mao, Yamazaki Tomoko, Imai Takayuki, Asada Yukinori, Matsuura Kazuto, Kawamura Sadafumi, Yamaguchi Kazunori, Yasuda Jun, Sugamura Kazuo, Katori Yukio, Satoh Kennichi, Tamai Keiichi
Division of Cancer Stem Cell, Miyagi Cancer Center Research Institute, Natori, Japan.
Department of Head and Neck Surgery, Miyagi Cancer Center, Natori, Japan.
Monoclon Antib Immunodiagn Immunother. 2020 Feb;39(1):6-11. doi: 10.1089/mab.2019.0040.
CD271 is a common receptor for all neurotrophins that is localized to neurons, endothelial cells, and the basal layer of the epithelium in normal tissue. Recently, we and others reported that CD271 plays essential roles in the development of squamous cell carcinoma, especially in tumor-initiating cells. Since little is known about how CD271 regulates cancer cell initiation and proliferation, antibodies that recognize different domains of CD271 are needed to enable investigation. Therefore, this study aimed to develop an antihuman CD271 antibody by immunizing mice with a CD271 antigen produced by a baculovirus. The antibody was named hCD271mAb#13, and it recognized cysteine-rich domain 1 with a higher affinity than the commercially available antibody ME20.4. We determined that hCD271mAb#13 is suitable for flow cytometry, Western blotting, immunocytochemistry, and immunohistochemistry of formalin-fixed paraffin-embedded tissue. Use of hCD271mAb#13 for CD271 labeling could enable detailed analyses of cancer cell regulation and other biological processes.
CD271是所有神经营养因子的共同受体，在正常组织中定位于神经元、内皮细胞和上皮基底层。最近，我们和其他研究人员报道，CD271在鳞状细胞癌的发展中起重要作用，特别是在肿瘤起始细胞中。由于对CD271如何调节癌细胞起始和增殖知之甚少，因此需要识别CD271不同结构域的抗体来进行研究。因此，本研究旨在通过用杆状病毒产生的CD271抗原免疫小鼠来开发一种抗人CD271抗体。该抗体被命名为hCD271mAb#13，它识别富含半胱氨酸的结构域1，其亲和力高于市售抗体ME20.4。我们确定hCD271mAb#13适用于流式细胞术、蛋白质印迹法、免疫细胞化学以及福尔马林固定石蜡包埋组织的免疫组织化学。使用hCD271mAb#13进行CD271标记能够对癌细胞调节和其他生物学过程进行详细分析。
