Dorfmueller Simone, Tan Hwee Ching, Ngoh Zi Xian, Toh Kai Yee, Peh Gary, Ang Heng-Pei, Seah Xin-Yi, Chin Angela, Choo Andre, Mehta Jodhbir S, Sun William
Experimental Therapeutics Centre, 31 Biopolis Way, Nanos level 3, Singapore, 138669 Singapore.
Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
Sci Rep. 2016 Feb 23;6:21661. doi: 10.1038/srep21661.
Cell surface antigens are important targets for monoclonal antibodies, but they are often difficult to work with due to their association with the cell membrane. Phage display is a versatile technique that can be applied to generate binders against difficult targets. Here we used antibody phage display to isolate a binder for a rare and specialized cell, the human corneal endothelial cell. The human corneal endothelium is a medically important cell layer; defects in this layer account for about half of all corneal transplants. Despite its importance, no specific antigens have been found to mark this cell type. By panning a phage library directly on human corneal endothelial cells, we isolated an antibody that bound to these cells and not the other types of corneal cells. Subsequently, we identified the antibody's putative target to be CD166 by immunoprecipitation and mass spectrometry. This approach can be used to isolate antibodies against other poorly-characterized cell types, such as stem cells or cancer cells, without any prior knowledge of their discriminating markers.
细胞表面抗原是单克隆抗体的重要靶点，但由于它们与细胞膜相关联，往往难以处理。噬菌体展示是一种通用技术，可用于生成针对难处理靶点的结合物。在这里，我们使用抗体噬菌体展示技术分离出一种针对罕见且特殊的细胞——人角膜内皮细胞的结合物。人角膜内皮是一层在医学上很重要的细胞层；该层的缺陷约占所有角膜移植的一半。尽管其很重要，但尚未发现特异性抗原可标记这种细胞类型。通过直接在人角膜内皮细胞上淘选噬菌体文库，我们分离出一种与这些细胞结合而不与其他类型角膜细胞结合的抗体。随后，我们通过免疫沉淀和质谱鉴定出该抗体的假定靶点为CD166。这种方法可用于分离针对其他特征不明确的细胞类型（如干细胞或癌细胞）的抗体，而无需事先了解它们的鉴别标志物。
