采用正常人和白血病患者的白细胞对人源噬菌体抗体库进行淘选,以获得对两种细胞表面蛋白特异的抗体.通过pⅧ展示系统,使抗体以多价展示于重组噬菌体颗粒表面,从上述两组中各挑选出48个克隆分别固定于环氧基片上,并以空白噬菌体和牛血清白蛋白作为对照,制成高密度噬菌体抗体芯片.取来自3名正常人和3名白血病患者的白细胞裂解物样品,用荧光染料Cy3标记,与噬菌体抗体芯片反应,对微阵共聚焦扫描得到的荧光图谱进行分析.在白血病白细胞表面蛋白的识别图谱中有8组斑点显著不同于正常图谱.由此表明,噬菌体抗体芯片可用于识别细胞表面蛋白.
A large human phage antibody library was subjected for panning with leukocytes from healthy donors and leukemia patients to select for specific antibodies against leukocytic surface proteins. Recombinant phage particles displaying multiple antibody fragments via pⅧ display system were used to prepare high-density phage antibody chips. Leukocyte lysate samples from 3 healthy donors and 3 leukemia patients were labeled with Cy3 fluorescence dye and reacted with the microarrays on the epoxy substrates. Eight sets of specific spots were found to have significantly distinct fluorescence intensities between the two groups of fluorescence profiles. It was demonstrated that phage antibody chips could be used for the recognition of cellular surface proteins.