细胞膜表面糖识别芯片的制备及其初步应用

目的制备可以对细胞膜表面糖链进行识别的凝集素芯片并进行初步应用。方法在芯片上制备 22 种凝集素点阵,提取组织和体液细胞进行荧光标记,利用凝集素与细胞膜表面糖链特异亲和性对细胞进行自动捕获,激光扫描仪和光镜检测。结果根据芯片上各个凝集素位点捕获细胞情况和该点凝集素的特异性获得细胞膜表面糖谱,并在光镜下对细胞数量形态进行观察。芯片的重复性和稳定性较好。结论该凝集素芯片可以对细胞进行自动捕获,对细胞膜表面糖链进行总体、即时的观察,对细胞膜糖链特征有一个总体的认识,建立各种细胞膜表面糖谱,为细胞膜表面糖链特征检测提供一个新的分析方法。

Preparation of Lectin Microarray and Cell Surface Glycome Profiling

Objective To develop a lectin microarray which can analyze the cell-surface glycome. Methods 22 lectins were printed on array,and the cells extracted from tissue and blood were labeled with fluorescence and caught by lectins through the distinctive binding specificities,and bound cells were directly detected by the laser fluorescence scanner or were observed under microscope after staining. Results Glycan profiling of cell membrane was obtained by distinctive binding specificities of lectins on microarray and the appearance of bound cells were observed under the microscope. The lectin micrearray was very stable and result was reproducible. Conclusion The lectin microarray can give integer and real-time analysis of the live cell-surface glycome without extraction and purification of glycoprotein,and establish the glycome profile of different cell surfaces. The technique provides a novel strategy for profiling global changes of the cell surface glycome.