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Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy
Li M(李密); Xiao XB(肖秀斌); Liu LQ(刘连庆); Xi N(席宁); Wang YC(王越超)
作者部门机器人学研究室
关键词Atomic Force Microscopy Peak Force Tapping Avidin-biotin Cd20 Rituximab Cell
发表期刊JOURNAL OF IMMUNOLOGICAL METHODS
ISSN0022-1759
2016
卷号436页码:41-49
收录类别SCI
WOS记录号WOS:000382346400006
产权排序1
资助机构National Natural Science Foundation of China [61503372, 61522312, 61327014, 61433017] ; Research Fund of the State Key Laboratory of Robotics [2014-Z07] ; CAS FEA International Partnership Program for Creative Research Teams
摘要Understanding the physicochemical properties of cell surface signalling molecules is important for us to uncover the underlying mechanisms that guide the cellular behaviors. Atomic force microscopy (AFM) has become a powerful tool for detecting the molecular interactions on individual cells with nanometer resolution. In this paper, AFM peak force tapping (PFT) imaging mode was applied to rapidly locate and visually map the CD20 molecules on human lymphoma cells using biochemically sensitive tips. First, avidin-biotin system was used to test the effectiveness of using PFT imaging mode to probe the specific molecular interactions. The adhesion images obtained on avidin-coated mica using biotin-tethered tips obviously showed the recognition spots which corresponded to the avidins in the simultaneously obtained topography images. The experiments confirmed the specificity and reproducibility of the recognition results. Then, the established procedure was applied to visualize the nanoscale organization of CD20s on the surface of human lymphoma Raji cells using rituximab (a monoclonal anti-CD20 antibody)-tethered tips. The experiments showed that the recognition spots in the adhesion images corresponded to the specific CD20-rituximab interactions. The cluster sizes of CD2Os on lymphoma Raji cells were quantitatively analyzed from the recognition images. Finally, under the guidance of fluorescence recognition, the established procedure was applied to cancer cells from a clinical lymphoma patient The results showed that there were significant differences between the adhesion images obtained on cancer cells and on normal cells (red blood cell). The CD20 distributions on ten cancer cells from the patient were quantified according to the adhesion images. The experimental results demonstrate the capability of applying PFT imaging to rapidly investigate the nanoscale biophysical properties of native membrane proteins on the cell surface, which is of potential significance in developing novel biomarkers for cancer diagnosis and drug development (C) 2016 Published by Elsevier B.V.
语种英语
WOS标题词Science & Technology ; Life Sciences & Biomedicine
WOS类目Biochemical Research Methods ; Immunology
关键词[WOS]CURVE-BASED AFM ; LIVING CELLS ; MOLECULAR RESOLUTION ; CD20 EXPRESSION ; NATIVE PROTEINS ; ROR1 ; SPECTROSCOPY ; ORGANIZATION ; RECEPTOR ; SURFACE
WOS研究方向Biochemistry & Molecular Biology ; Immunology
引用统计
文献类型期刊论文
条目标识符http://ir.sia.cn/handle/173321/19202
专题机器人学研究室
通讯作者Liu LQ(刘连庆); Xi N(席宁)
作者单位1.State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016, China
2.University of Chinese Academy of Sciences, Beijing 100049, China
3.Department of Lymphoma, Affiliated Hospital of Military Medical Academy of Sciences, Beijing 100071, China
4.Department of Electrical and Computer Engineering, Michigan State University, East Lansing, MI 48824, USA
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GB/T 7714
Li M,Xiao XB,Liu LQ,et al. Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy[J]. JOURNAL OF IMMUNOLOGICAL METHODS,2016,436:41-49.
APA Li M,Xiao XB,Liu LQ,Xi N,&Wang YC.(2016).Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy.JOURNAL OF IMMUNOLOGICAL METHODS,436,41-49.
MLA Li M,et al."Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy".JOURNAL OF IMMUNOLOGICAL METHODS 436(2016):41-49.
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