Liu Jiangnan, Yi Bin, Zhang Zhe, Cao Yi
Laboratory of Molecular and Experimental Pathology, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, 650223, China.
Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming, 650223, China.
Front Med. 2016 Jun;10(2):204-11. doi: 10.1007/s11684-016-0443-1. Epub 2016 Apr 18.
CD176 (Thomsen-Friedenreich antigen) is a tumor-associated carbohydrate epitope (glycotope) functionally involved in blood spread and liver metastasis of cancer cells by mediating the adhesion of cancer cells to endothelial cells and hepatocytes, respectively. CD176 could be a promising target for antitumor immunotherapy. We applied B lymphocytes obtained from mice immunized with CD176 antigen and constructed a phage display library. A positive clone of CD176 single-chain variable antibody fragment (scFv) was successfully screened from this library. The CD176 scFv was expressed in Escherichia coli and purified. The purified scFv can bind to the natural CD176 expressed on the surface of cancer cells. Furthermore, the CD176 scFv inhibits the adhesion of CD176(+) cancer cells to endothelial cells and hepatocytes. This CD176 scFv provides a basis for future development of recombinant CD176-specific antibodies that can be used in therapeutic application.
CD176（汤姆森-弗里德赖希抗原）是一种肿瘤相关碳水化合物表位（糖表位），通过分别介导癌细胞与内皮细胞和肝细胞的黏附，在癌细胞的血行播散和肝转移中发挥功能作用。CD176可能是抗肿瘤免疫治疗的一个有前景的靶点。我们应用从用CD176抗原免疫的小鼠获得的B淋巴细胞，构建了一个噬菌体展示文库。从该文库中成功筛选出CD176单链可变抗体片段（scFv）的一个阳性克隆。CD176 scFv在大肠杆菌中表达并纯化。纯化后的scFv能与癌细胞表面表达的天然CD176结合。此外，CD176 scFv可抑制CD176(+)癌细胞与内皮细胞和肝细胞的黏附。这种CD176 scFv为未来开发可用于治疗应用的重组CD176特异性抗体奠定了基础。
