物理化学学报 >> 2006, Vol. 22 >> Issue (07): 777-779.doi: 10.3866/PKU.WHXB20060702

研究论文 上一篇    下一篇

高密度噬菌体抗体芯片对细胞表面蛋白的识别

岑晓东;王文娟;赵新生;汪玄;沈悌;谭涛超;毕群;朱圣庚   

  1. 北京大学生命科学学院, 北京 100871; 北京大学化学与分子工程学院化学生物学系, 分子动态与稳态结构国家重点实验室, 北京 100871; 北京协和医院, 北京 100730
  • 收稿日期:2005-12-29 修回日期:2006-02-23 发布日期:2006-06-27
  • 通讯作者: 赵新生;朱圣庚 E-mail:zhusg@pku.edu.cn;zhaoxs@pku.edu.cn

Recognition of Cell Surface Proteins by High-density Phage Antibody Chip

CEN Xiao-Dong;WANG Wen-Juan;ZHAO Xin-Sheng;WANG Xuan;SHEN Ti;TAN Tao-Chao;BI Qun;ZHU Sheng-Geng   

  1. College of Life Sciences, Peking University, Beijing 100871, P. R. China; State Key Laboratory for Structural Chemistry of Unstable and Stable Species, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Peking University, Beijing 100871, P. R. China; Peking Union Medical College Hospital, Beijing 100730, P. R. China
  • Received:2005-12-29 Revised:2006-02-23 Published:2006-06-27
  • Contact: ZHAO Xin-Sheng;ZHU Sheng-Geng E-mail:zhusg@pku.edu.cn;zhaoxs@pku.edu.cn

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摘要： 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.

关键词: 噬菌体抗体库, pVIII展示系统, 高密度噬菌体抗体芯片, 白血病, 白细胞表面蛋白

Abstract: 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.

Key words: Phage antibody library, pVIII display system, High-density phage antibody chip, Leukemia, Leukocytic surface protein