Detecting CD20-Rituximab interaction forces using AFM single-molecule force spectroscopy  

作　　者：LI Mi LIU LianQing XI Ning WANG YueChao DONG ZaiLi LI GuangYong XIAO XiuBin ZHANG WeiJing 

机构地区：[1]State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016, China [2]Graduate University of Chinese Academy of Sciences, Beijing 100049, China [3]Department of Electrical and Computer Engineering, Michigan State University, East Lansing 48824, USA [4]Department of Electrical and Computer Engineering, University of Pittsburgh, Pittsburgh 15261, USA [5]Department of Lymphoma, Affiliated Hospital of Military Medical Academy of Sciences, Beijing 100071, China

出　　处：《Chinese Science Bulletin》2011年第35期3829-3835,共7页

基　　金：supported by the National Natural Science Foundation of China (60904095);National High Technology Research and Development Program of China (2009AAO3Z316);CAS FEA International Partnership Program for Creative Research Teams

摘　　要：The invention of atomic force microscopy(AFM) has provided new technology for measuring specific molecular interaction forces.Using AFM single-molecule force spectroscopy(SMFS) techniques,CD20-Rimximab rupture forces were measured on purified CD20 proteins,Raji cells,and lymphoma patient B cells.Rimximab molecules were linked onto AFM tips using AFM probe functionalization technology,and purified CD20 proteins were attached to mica using substrate functionalization technology.Raji cells(a lymphoma cell line) or lymphoma patient cells were immobilized on a glass substrate via electrostatic adsorption and chemical fixation.The topography of the purified CD20 proteins,Raji cells,and patient lymphoma cells was visualized using AFM imaging and the differences in the rupture forces were analyzed and measured.The results showed that the rupture forces between the CD20 proteins on Raji cells and Rituximab were markedly smaller than those for purified CD20 proteins and CD20 proteins on lymphoma patient B cells.These findings provide an effective experimental method for investigating the mechanisms underlying the variable efficacy of Rituximab.The invention of atomic force microscopy （AFM） has provided new technology for measuring specific molecular interaction forces. Using AFM single-molecule force spectroscopy （SMFS） techniques, CD20-Rituximab rupture forces were measured on purified CD20 proteins, Raji cells, and lymphoma patient B cells. Rituximab molecules were linked onto AFM tips using AFM probe functionalization technology, and purified CD20 proteins were attached to mica using substrate functionalization technology. Raji cells （a lymphoma cell line） or lymphoma patient cells were immobilized on a glass substrate via electrostatic adsorption and chemical fixation. The topography of the purified CD20 proteins, Raji cells, and patient lymphoma cells was visualized using AFM imaging and the differences in the rupture forces were analyzed and measured. The results showed that the rupture forces between the CD20 proteins on Raji cells and Rituximab were markedly smaller than those for purified CD20 proteins and CD20 proteins on lymphoma patient B cells. These findings provide an effective experimental method for investigating the mechanisms underlying the variable efficacy of Rituximab.

关 键 词：原子力显微镜 相互作用力 单分子 光谱 RAJI细胞 蛋白质纯化 功能化技术 AFM探针 

分 类 号：R733.1[医药卫生—肿瘤]