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题名:
Rapid recognition and functional analysis of membrane proteins on human cancer cells using atomic force microscopy
作者: 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
通讯作者: 刘连庆 ; 席宁
关键词: Atomic force microscopy ; Peak force tapping ; Avidin-biotin ; CD20 ; Rituximab ; Cell
刊名: JOURNAL OF IMMUNOLOGICAL METHODS
ISSN号: 0022-1759
出版日期: 2016
卷号: 436, 页码:41-49
收录类别: SCI
产权排序: 1
项目资助者: 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记录号: WOS:000382346400006
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods ; Immunology
关键词[WOS]: CURVE-BASED AFM ; LIVING CELLS ; MOLECULAR RESOLUTION ; CD20 EXPRESSION ; NATIVE PROTEINS ; ROR1 ; SPECTROSCOPY ; ORGANIZATION ; RECEPTOR ; SURFACE
研究领域[WOS]: Biochemistry & Molecular Biology ; Immunology
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内容类型: 期刊论文
URI标识: http://ir.sia.cn/handle/173321/19202
Appears in Collections:机器人学研究室_期刊论文

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Recommended Citation:
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,2016-01-01,436:41-49.
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