Son Yeon Sung, Hong Hyo Jeong
Therapeutic Antibody Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejon, Republic of Korea.
J Microbiol. 2007 Dec;45(6):547-52.
Previously, we generated monoclonal antibodies (MAbs) that bound to the surface of human embryonic stem cells (hESCs) in an attempt to discover new hESC-specific surface markers. In this study, MAb 47-235 (IgG1, kappa) was selected for further characterization. The MAb bound to the surface of undifferentiated hESCs but did not bind to mouse ESCs or mouse embryonic fibroblast cells in flow cytometric analysis. The antibody immunoprecipitated a 47 kDa protein from the lysates of cell surface-biotinylated hESCs. Identification of the protein by quadrupole time of flight tandem mass spectrometry revealed that 47-235 binds to Ag 243-5 protein of Mycoplasma arginini. BM-Cyclin treatment of the hESCs that reacted with 47-235 resulted in loss of mycoplasma DNA and the reactivity to 47-235. Nevertheless, the hESCs that were reactive to 47-235 maintained self-renewal and pluripotency and thus could be differentiated into three embryonic germ layers.
此前，我们制备了与人类胚胎干细胞（hESC）表面结合的单克隆抗体（MAb），试图发现新的hESC特异性表面标志物。在本研究中，选择了单克隆抗体47-235（IgG1，κ）进行进一步表征。在流式细胞术分析中，该单克隆抗体与未分化的hESC表面结合，但不与小鼠胚胎干细胞或小鼠胚胎成纤维细胞结合。该抗体从细胞表面生物素化的hESC裂解物中免疫沉淀出一种47 kDa的蛋白质。通过四极杆飞行时间串联质谱对该蛋白质进行鉴定，结果显示47-235与精氨酸支原体的Ag 243-5蛋白结合。用BM-Cyclin处理与47-235反应的hESC，导致支原体DNA丢失以及对47-235的反应性丧失。然而，对47-235有反应的hESC保持了自我更新和多能性，因此可以分化为三个胚胎胚层。
